Pyrimidine compounds

ABSTRACT

This invention relates to a method for treating inflammatory diseases or immune diseases, developmental or degenerative diseases, or tissue injuries. The method includes administering to a subject in need thereof an effective amount of one or more compounds of formula (I). Each variable in this formula is defined in the specification.

CROSS REFERENCE TO RELATED APPLICATIONS

Pursuant to 35 USC § 119(e), this application claims priority to U.S.Provisional Application Ser. No. 60/690,267, filed Jun. 14, 2005, andU.S. Provisional Application Ser. No. 60/798,596, filed May 8, 2006, thecontents of which are incorporated herein by reference.

BACKGROUND

Chemokines are a family of cytokines that regulate the adhesion andtransendothelial migration of leukocytes during an immune orinflammatory reaction (Mackay C. R., Nat. Immunol., (2001) 2:95; Olsonet al., Am. J. Physiol. Regul. Integr. Comp. Physiol., (2002) 283:R7).Chemokines also regulate T cells and B cells trafficking and homing, andcontribute to the development of lymphopoietic and hematopoietic systems(Ajuebor et al., Biochem. Pharmacol., (2002) 63:1191). Approximately 50chemokines have been identified in humans. They can be classified into 4subfamilies, i.e., CXC, CX3C, CC, and C chemokines, based on thepositions of the conserved cysteine residues at the N-terminal (Onufferet al., Trends Pharmacol Sci., (2002) 23:459). The biological functionsof chemokines are mediated by their binding and activation of Gprotein-coupled receptors (GPCRs) on the cell surface. Take CXCR4receptor for example, it can be activated by Stromal-derived factor-1 orSDF-1, a member of CXC chemokines.

SDF-1 was originally cloned from bone marrow stromal cell lines andfound to act as a growth factor for progenitor B cells (Nishikawa etal., Eur. J. Immunol., (1988) 18:1767). SDF-1 also induces bone marrowcolonization of hematopoietic precursor cells during embryogenesis(Bleul et al., J. Exp. Med., (1996) 184:1101). The physiologicalfunction of SDF-1 is mediated by CXCR4 receptor. Mice lacking SDF-1 orCXCR4 receptor show lethal abnormality in bone marrow myelopoiesis, Bcell lymphopoiesis, and cerebellar development (Nagasawa et al., Nature,(1996) 382:635; Ma et al., Proc. Natl. Acad. Sci., (1998) 95:9448; Zouet al., Nature (1998) 393:595; Lu et al., Proc. Natl. Acad. Sci. (2002)99:7090). CXCR4 receptor is expressed broadly in a variety of tissues,particularly in immune and central nervous systems, and has beendescribed as the major co-receptor for HIV-1/2 on T lymphocytes.Although initial interest in CXCR4 antagonism focused on its potentialapplication to AIDS treatment (Bleul et al., Nature (1996) 382:829), itis now becoming clear that CXCR4 receptor and SDF-1 are also involved inother pathological conditions such as rheumatoid arthritis, asthma, andtumor metastases (Buckley et al., J. Immunol., (2000) 165:3423). CXCR4receptor and SDF-1 are also found widely expressed in many tissuesduring embryonic development. Further, the CXCR4/SDF-1 pathway has beenshown to be critically involved in the regeneration of several tissueinjury models. Specifically, it has been found that the SDF-1 level iselevated at an injured site and CXCR4-positive cells activelyparticipate in the tissue regenerating process.

SUMMARY

This invention is based on the discovery that certain pyrimidinecompounds are effective in treating inflammatory and immune diseases(e.g., retinopathy), developmental or degenerative diseases, or tissueinjuries through their binding to chemokine receptors (e.g., CXCR3 orCXCR4 receptors). In addition, these compounds, when used in combinationwith G-CSF growth factor, exhibited synergistic effects in stem cellsand endothelial progenitor cells mobilization.

In one aspect, this invention features pyrimidine compounds of formula(I) and their salts:

In this formula, X is —N(R_(a))— or —O—; or X, together with R₅, isC₃-C₂₀ heterocycloalkyl; or X, together with L₂ and L₃, is C₃-C₂₀heterocycloalkyl; each of L₁ and L₂, independently, is C₁-C₁₀ alkylene,C₁-C₁₀ heteroalkylene, —C(O)—, or deleted; or L₁, together with L₃, R₄,and the nitrogen attached to R₄, is C₃-C₂₀ heterocycloalkyl, or L₂,together with L₃ and X, is C₃-C₂₀ heterocycloalkyl; L₃ is —N(R_(b))—,—O—, aryl, heteroaryl, or C₃-C₂₀ cycloalkyl; or L₃, together with L₁,R₄, and the nitrogen attached to R₄, is C₃-C₂₀ heterocycloalkyl; or L₃,together with L₂ and X, is C₃-C₂₀ heterocycloalkyl; each of R₁, R₂, andR₃, independently, is H, C₁-C₁₀ alkyl, C₃-C₂₀ cycloalkyl, C₃-C₂₀heterocycloalkyl, aryl, heteroaryl, halo, CN, OR_(c), COOR_(c),OC(O)R_(c), C(O)R_(c), C(O)NR_(c)R_(d), or NR_(c)R_(d); R₄ is H, C₁-C₁₀alkyl, C₃-C₂₀ cycloalkyl, C₃-C₂₀ heterocycloalkyl, aryl, or heteroaryl;and R₅ is C₃-C₂₀ cycloalkyl, C₃-C₂₀ heterocycloalkyl, aryl, heteroaryl,or C₁-C₁₀ alkyl substituted with C₃-C₂₀ cycloalkyl, C₃-C₂₀heterocycloalkyl, or N(R_(e)R_(f)); or R₅, together with X, is C₃-C₂₀heterocycloalkyl; in which each of R_(a), R_(b), R_(c), R_(d), R_(e),and R_(f), independently, is H, C₁-C₁₀ alkyl, C₃-C₂₀ cycloalkyl, C₃-C₂₀heterocycloalkyl, aryl, heteroaryl, or —C(O)R; R being H, C₁-C₁₀ alkyl,C₃-C₂₀ cycloalkyl, C₃-C₂₀ heterocycloalkyl, aryl, or heteroaryl.

Referring to formula (I), a subset of the pyrimidine compounds describedabove are those in which X can be —N(R_(a))—, each of L₁ and L₂,independently, can be C₁-C₁₀ alkylene, —C(O)—, or deleted; and L₃ can be—N(R_(b))—, —O—, aryl, or C₃-C₂₀ cycloalkyl. As one example, in certainpyrimidine compounds, L₃ is —N(R_(b))—. In these compounds, R₅ can beC₃-C₂₀ cycloalkyl (e.g.,

C₁-C₁₀ alkyl substituted with C₃-C₂₀ cycloalkyl (e.g.,

or C₃-C₂₀ heterocycloalkyl substituted with C₁-C₁₀ alkyl (e.g.

R_(b) can be C₁-C₁₀ alkyl substituted with N(R′R″), in which each of R′and R″, independently, is H, C₁-C₁₀ alkyl, C₃-C₂₀ cycloalkyl, C₃-C₂₀heterocycloalkyl, aryl, or heteroaryl; and R₁ can be C₃-C₂₀heterocycloalkyl (e.g.,

As another example, in certain pyrimidine compounds, L₃ can be aryl(e.g., phenylene). In these compounds, R₅ can be C₁-C₁₀ alkylsubstituted with C₃-C₂₀ heterocycloalkyl (e.g.

or

or C₁-C₁₀ alkyl substituted with N(R_(e)R_(f)) (e.g.,

or R₅, together with X, is C₃-C₂₀ heterocycloalkyl (e.g.,

R₃ can be H, halo C₁-C₁₀ alkyl, OR_(c), NR_(c)R_(d), or C₃-C₂₀heterocycloalkyl optionally substituted with C₁-C₁₀ alkyl, C₃-C₂₀cycloalkyl, C₃-C₂₀ heterocycloalkyl, aryl, OR′, C(O)R′, COOR′,C(O)N(R′R″), SO₂R′, C(S)N(R′R″), OSO₃R′, or PO(OR′)₂, in which each ofR′ and R″, independently, is H, C₁-C₁₀ alkyl, C₃-C₂₀ cycloalkyl, C₃-C₂₀heterocycloalkyl, aryl, or heteroaryl. For example, R₃ can be H, Cl,CH₃, OPh,

optionally substituted with OH,

optionally substituted with C₁-C₁₀ alkyl, C₃-C₂₀ cycloalkyl, C₃-C₂₀heterocycloalkyl, aryl, OH, C(O)R′, COOR′, C(O)N(R′R″), SO₂R′,C(S)N(R′R″), OSO₃R′, PO(OR′)₂, or NH(R′) substituted with OH orNHC(O)R″. As another example, in certain pyrimidine compounds, L₃ can beC₃-C₂₀ cycloalkyl (e.g., cyclohexylene). In these compounds, R₅ can beC₁-C₁₀ alkyl substituted with N(R_(e)R_(f)) (e.g.,

and R₃ can be C₃-C₂₀ heterocycloalkyl substituted with C₁-C₁₀ alkyl,C₃-C₂₀ cycloalkyl, C₃-C₂₀ heterocycloalkyl, aryl, OR′, C(O)R′, COOR′,C(O)N(R′R″), SO₂R′, or C(S)N(R′R″), in which each of R′ and R″,independently, is H, C₁-C₁₀ alkyl, C₃-C₂₀ cycloalkyl, C₃-C₂₀heterocycloalkyl, aryl, or heteroaryl.

The term “alkyl” refers to a saturated or unsaturated, linear orbranched hydrocarbon moiety, such as —CH₃, —CH₂—CH═CH₂, or branched—C₃H₇. The term “alkylene” refers to a divalent, saturated orunsaturated, linear or branched hydrocarbon moiety, such as —CH₂— or—CH═CH—. The term “heteroalkylene” refers to an alkylene moiety havingat least one heteroatom (e.g., N, O, or S). The term “cycloalkyl” refersto a saturated or unsaturated, non-aromatic, cyclic hydrocarbon moiety,such as cyclohexyl or cyclohexen-3-yl. The term “heterocycloalkyl”refers to a saturated or unsaturated, non-aromatic, cyclic moiety havingat least one ring heteroatom (e.g., N, O, or S), such as4-tetrahydropyranyl or 4-pyranyl. The term “aryl” refers to ahydrocarbon moiety having one or more aromatic rings. Examples of arylmoieties include phenyl (Ph), phenylene, naphthyl, naphthylene, pyrenyl,anthryl, and phenanthryl. The term “heteroaryl” refers to a moietyhaving one or more aromatic rings that contain at least one heteroatom(e.g., N, O, or S). Examples of heteroaryl moieties include furyl,furylene, fluorenyl, pyrrolyl, thienyl, oxazolyl, imidazolyl, thiazolyl,pyridyl, pyrimidinyl, quinazolinyl, quinolyl, isoquinolyl and indolyl.

Alkyl, alkylene, heteroalkylene, cycloalkyl, heterocycloalkyl, aryl, andheteroaryl mentioned herein include both substituted and unsubstitutedmoieties, unless specified otherwise. Possible substituents oncycloalkyl, heterocycloalkyl, aryl, and heteroaryl include, but are notlimited to, C₁-C₁₀ alkyl, C₂-C₁₀ alkenyl, C₂-C₁₀ alkynyl, C₃-C₂₀cycloalkyl, C₃-C₂₀ cycloalkenyl, C₃-C₂₀ heterocycloalkyl, C₃-C₂₀heterocycloalkenyl, C₁-C₁₀ alkoxy, aryl, aryloxy, heteroaryl,heteroaryloxy, amino, C₁-C₁₀ alkylamino, C₁-C₂₀ dialkylamino, arylamino,diarylamino, hydroxyl, halogen, thio, C₁-C₁₀ alkylthio, arylthio, C₁-C₁₀alkylsulfonyl, arylsulfonyl, acylamino, aminoacyl, aminothioacyl,amidino, guanidine, ureido, cyano, nitro, acyl, thioacyl, acyloxy,carboxyl, and carboxylic ester. On the other hand, possible substituentson alkyl, alkylene, or heteroalkylene include all of the above-recitedsubstituents except C₁-C₁₀ alkyl, C₂-C₁₀ alkenyl, and C₂-C₁₀ alkynyl.Cycloalkyl, heterocycloalkyl, aryl, and heteroaryl can also be fusedwith each other.

In another aspect, this invention features pyrimidine compounds offormula (I) shown above in which X is —N(R_(a))— or —O—; each of L₁ andL₂, independently, is C₁-C₁₀ alkylene, C₁-C₁₀ heteroalkylene, —C(O)—, ordeleted; L₃ is —N(R_(b))—, C₃-C₂₀ cycloalkyl, aryl, heteroaryl, ordeleted; R₁ is H, C₁-C₁₀ alkyl, C₃-C₂₀ cycloalkyl, aryl, heteroaryl,halo, CN, OR_(c), COOR_(c), OC(O)R_(c), C(O)R_(c), C(O)NR_(c)R_(d), orNR_(c)R_(d); each of R₂ and R₃, independently, is H, C₁-C₁₀ alkyl,C₃-C₂₀ cycloalkyl, C₃-C₂₀ heterocycloalkyl, aryl, heteroaryl, halo, CN,OR_(e), COOR_(e), OC(O)R_(e), C(O)R_(e), C(O)NR_(e)R_(f), orNR_(e)R_(f); and each of R₄ and R₅, independently, is H, C₁-C₁₀ alkyl,C₃-C₂₀ cycloalkyl, C₃-C₂₀ heterocycloalkyl, aryl, or heteroaryl; or R₄and R₅ together are C₁-C₁₀ alkylene or C₁-C₁₀ heteroalkylene; in whicheach of R_(a), R_(b), R_(c), R_(d), R_(e), and R_(f), independently, isH, C₁-C₁₀ alkyl, C₃-C₂₀ cycloalkyl, C₃-C₂₀ heterocycloalkyl, aryl, orheteroaryl; or a salt thereof.

Referring to formula (I), a subset of the just-described pyrimidinecompounds are those in which X is —N(R_(a))—; each of L₁ and L₂,independently, is C₁-C₁₀ alkylene; L₃ is deleted; R₁ is NR_(c)R_(d);each of R₂ and R₃, independently, is H, C₁-C₁₀ alkyl, halo, or C₃-C₂₀cycloalkyl; and each of R₄ and R₅, independently, is H or C₃-C₂₀cycloalkyl; or R₄ and R₅ together are C₁-C₁₀ alkylene or C₁-C₁₀heteroalkylene. In these compounds, R₅ can be

or R₄ and R₅ together can be —CH₂CH₂—; one of R_(c) and R_(d) can beC₁-C₁₀ alkyl substituted with N(RR′) or aryl, in which each of R and R′,independently, is H, C₁-C₁₀ alkyl, C₃-C₂₀ cycloalkyl, C₃-C₂₀heterocycloalkyl, aryl, or heteroaryl (e.g., one of

substituted with C₁-C₁₀ alkyl, which is in turn substituted withC₃-C₂₀-heterocycloalkyl or OR, R being H, C₁-C₁₀ alkyl, C₃-C₂₀cycloalkyl, C₃-C₂₀ heterocycloalkyl, aryl, or heteroaryl.

In still another aspect, this invention features a method for treatingan inflammatory or immune disease, a developmental or degenerativedisease, or a tissue injury. The method includes administering to asubject in need thereof an effective amount of one or more pyrimidinecompounds of formula (I) shown above.

The term “treating” or “treatment” refers to administering one or morepyrimidine compounds to a subject, who has an above-described disease, asymptom of such a disease, or a predisposition toward such a disease,with the purpose to confer a therapeutic effect, e.g., to cure, relieve,alter, affect, ameliorate, or prevent the above-described disease, thesymptom of it, or the predisposition toward it.

An inflammatory disease is characterized by a local or systemic, acuteor chronic inflammation. Examples include retinopathy, inflammatorydermatoses (e.g., dermatitis, eczema, atopic dermatitis, allergiccontact dermatitis, urticaria, necrotizing vasculitis, cutaneousvasculitis, hypersensitivity vasculitis, eosinophilic myositis,polymyositis, dermatomyositis, and eosinophilic fasciitis), inflammatorybowel diseases (e.g., Crohn's disease and ulcerative colitis),hypersensitivity lung diseases (e.g., hypersensitivity pneumonitis,eosinophilic pneumonia, delayed-type hypersensitivity, interstitial lungdisease or ILD, idiopathic pulmonary fibrosis, and ILD associated withrheumatoid arthritis), asthma, and allergic rhinitis.

An immune disease is characterized by a hyper- or hypo-reaction of theimmune system. Examples include autoimmune diseases (e.g., rheumatoidarthritis, psoriatic arthritis, systemic lupus erythematosus, myastheniagravis, juvenile onset diabetes, glomerulonephritis, autoimmunethroiditis, ankylosing spondylitis, systemic sclerosis, and multiplesclerosis), acute and chronic inflammatory diseases (e.g., systemicanaphylaxia or hypersensitivity responses, drug allergies, insect stingallergies, graft rejection, including allograft rejection, andgraft-versus-host disease), Sjogren's syndrome, human immunodeficiencyvirus infection, cancer (e.g., brain, breast, prostate, colon, kidney,ovary, thyroid, lung, and haematopoietic cancer), and tumor metastasis.

Developmental diseases are growth or differentiation related disordersthat lead to loss-of-function or gain-of-function. Degenerative diseasesgenerally refer to change of a tissue to a lower or less functionalform. Examples of a developmental or degenerative disease include spinalmuscular atrophy, Duchenne muscular dystrophy, Parkinson's disease, andAlzheimer's disease. Tissue injuries can be caused by oxidative stress(e.g., ischemia-reperfusion in stroke or myocardial infarction),complement activation, graft rejection, chemicals (e.g., alcohol-inducedliver damage or mucosal tissue injuries in cancer therapy), viralinfection (e.g., glomerular injuries associated with hepatitis Cinfection), and mechanical forces (e.g., sports injury). Examples oftissue injuries include brain injury, heart injury, liver damage,skeletal muscle injury, kidney damage, pancreatic injury, lung injury,skin injury, and gastrointestinal tract injury.

A subject in need of treatment of an above-described disease can also beconcurrently administered with a pyrimidine compound described above andone or more other therapeutic agents. Examples of such a therapeuticagent include a G-CSF growth factor, a steroidal or a non-steroidalanti-inflammatory drug, a COX2 inhibitor, a leukotriene receptorinhibitor, a prostaglandin modulator, a TNF modulator, and animmunosuppressive agent (e.g., cyclosporine A). The term “concurrentlyadministered” refers to administering a pyrimidine compound and one ormore other therapeutic agents at the same time or at different timesduring the period of treatment.

In still another aspect, this invention features a method for enhancingmigration of bone marrow-derived cells to blood. The method includesadministering to a subject in need thereof an effective amount of one ormore pyrimidine compounds of formula (I) shown above. The term “bonemarrow-derived cells” refers to cells originating from bone marrow.Examples of bone marrow-derived cells include, but are not limited to,CD34+ cells and CD133+ cells. Preferrably, bone marrow-derived cells arestem cells or endothelial progenitor cells.

In still another aspect, this invention features a chemical syntheticmethod. The method includes reacting a compound of the formula

with a compound of the formula

to give a compound of formula (II):

In this formula, A is phenylene, cyclohexylene,

each of R₁, R₂, and R₃, independently, is H, C₁-C₁₀ alkyl, C₃-C₂₀cycloalkyl, C₃-C₂₀ heterocycloalkyl, aryl, heteroaryl, halo, CN, OR_(a),COOR_(a), OC(O)R_(a), C(O)R_(a), C(O)NR_(a)R_(b), or NR_(a)R_(b); R₆ ishalo; and R₇ is a amino-protecting group (e.g., t-butoxycarbonyl,benzyloxycarbonyl, acetyl, phenylcarbonyl, or trialkylsilyl); in whicheach of R_(a) and R_(b), independently, is H, C₁-C₁₀ alkyl, C₃-C₂₀cycloalkyl, C₃-C₂₀ heterocycloalkyl, aryl, heteroaryl, or —C(O)R; Rbeing H, C₁-C₁₀ alkyl, C₃-C₂₀ cycloalkyl, C₃-C₂₀ heterocycloalkyl, aryl,or heteroaryl.

The method can further include deprotecting the compound of formula (II)to give a compound of formula (III):

(III). In a subset of compounds of formula (III), R₁ is N(R_(a)R_(b)),in which R_(a) and R_(b), independently, is H, C₁-C₁₀ alkyl, C₃-C₂₀cycloalkyl, C₃-C₂₀ heterocycloalkyl, aryl, or heteroaryl. In thesecompounds, one of R_(a) and R_(b) can be

In another subset of compounds of formula (III), R₃ is

optionally substituted with C₁-C₁₀ alkyl, C₃-C₂₀ cycloalkyl, C₃-C₂₀heterocycloalkyl, or aryl.

Referring to formula (II), when R₃ is halo, a compound of formula (II)can further react with a compound of the formula

to give a compound of formula (IV):

(IV). In this formula, Y is —O—, —CH₂—, or —N(R_(c))—, in which R_(c) isH, C₁-C₁₀ alkyl, C₃-C₂₀ cycloalkyl, C₃-C₂₀ heterocycloalkyl, aryl,heteroaryl, or halo. The compound of formula (IV) can be deprotected togive a compound of formula (V):

Referring to formula (IV), when Y is NH, a compound of formula (IV) canfurther react with a compound of the formula

to give a compound of formula (VI):

(VI), in which Z is —CH₂— or —C(O)—; R_(d) is C₁-C₁₀ alkyl, C₃-C₂₀cycloalkyl, C₃-C₂₀ heterocycloalkyl, aryl, or heteroaryl; and R_(e) ishalo. The compound of formula (VI) can be deprotected to give a compoundof formula (VII):

As another example, when Y is NH, a compound of formula (IV) can furtherreact with a compound of the formula

to give an imine compound, followed by reducing the imine compound togive a compound of formula (VI). In the formula

R_(d) is C₁-C₁₀ alkyl, C₃-C₂₀ cycloalkyl, C₃-C₂₀ heterocycloalkyl, aryl,or heteroaryl; and R_(e) is H. As another example, when Y is NH, acompound of formula (IV) can further react with a compound of theformula

to give an ester compound, followed by hydrolyzing the ester compound togive a compound of formula (VIII):

(VIII), in which R_(d) is C₁-C₁₀ alkyl, C₃-C₂₀ cycloalkyl, C₃-C₂₀heterocycloalkyl, aryl, or heteroaryl. The compound of formula (VIII)can be deprotected to give a compound of formula (IX):

(IX). As a further example, when Y is NH, a compound of formula (IV) canfurther react with a compound of the formula

to give a compound of formula (X):

(X), in which R_(d) is C₁-C₁₀ alkyl, C₃-C₂₀ cycloalkyl, C₃-C₂₀heterocycloalkyl, aryl, or heteroaryl. The compound of formula (X) canbe deprotected to give a deprotected compound, followed by hydrolyzingthe deprotected compound to give a compound of formula (XI):

Referring to formula (II), when R₃ is halo, a compound of formula (II)can further react with a compound of the formula R_(c)OH to give acompound of formula (XII):

in which R_(c) is C₁-C₁₀ alkyl, C₃-C₂₀ cycloalkyl, C₃-C₂₀heterocycloalkyl, aryl, or heteroaryl. The compound of formula (XII) canbe deprotected to give a compound of formula (XIII):

Referring to formula (II), when R₂ is CN, the compound of formula (II)can be reduced to give a compound of formula (XIV):

The compound of formula (XIV) can further react with a compound of theformula R_(c)NH₂ to give an imine compound, followed by reducing theimine compound to give a compound of formula (XV):

in which R_(c) is C₁-C₁₀ alkyl, C₃-C₂₀ cycloalkyl, C₃-C₂₀heterocycloalkyl, aryl, or heteroaryl. The compound of formula (XV) canbe deprotected to give a compound of formula (XVI):

In a further aspect, the invention features a chemical synthetic methodthat includes reacting a compound of the formula

with a compound of the formula

to give a compound of formula (XVII):

In formula (XVII), each of R₁ and R₂, independently, is H, C₁-C₁₀ alkyl,C₃-C₂₀ cycloalkyl, C₃-C₂₀ heterocycloalkyl, aryl, heteroaryl, halo, CN,OR_(a), COOR_(a), OC(O)R_(a), C(O)R_(a), C(O)NR_(a)R_(b), orNR_(a)R_(b); R₃ is halo; and R₆ is a amino-protecting group; in whicheach of R_(a) and R_(b), independently, is H, C₁-C₁₀ alkyl, C₃-C₂₀cycloalkyl, C₃-C₂₀ heterocycloalkyl, aryl, heteroaryl, or —C(O)R; Rbeing H, C₁-C₁₀ alkyl, C₃-C₂₀ cycloalkyl, C₃-C₂₀ heterocycloalkyl, aryl,or heteroaryl. The method can further include protecting the compound offormula (XX), followed by reacting the protected compound of formula(XVII) with a compound of the formula

to give a compound of formula (XVIII):

In formula (XVIII), R₇ is a amino-protecting group; and Y is —O—, —CH₂—,or —N(R_(c))—, in which R_(c) is H, C₁-C₁₀ alkyl, C₃-C₂₀ cycloalkyl,C₃-C₂₀ heterocycloalkyl, aryl, heteroaryl, or halo.

Referring to formula (XVIII), when Y is NH, the method can furtherinclude: (1) reacting the compound of formula (XVIII) with a compound ofthe formula

to give an ester compound; (2) hydrolyzing the ester compound to give anacid compound; and (3) deprotecting the acid compound to give a compoundof formula (XIX):

(XIX), in which R_(d) is C₁-C₁₀ alkyl, C₃-C₂₀ cycloalkyl, C₃-C₂₀heterocycloalkyl, aryl, or heteroaryl; and R_(e) is halo.

In addition, this invention encompasses a pharmaceutical compositionthat contains an effective amount of at least one of the above-mentionedpyrimidine compounds and a pharmaceutically acceptable carrier.

The pyrimidine compounds described above include the compoundsthemselves, as well as their salts, prodrugs, and solvates, ifapplicable. A salt, for example, can be formed between an anion and apositively charged group (e.g., amino) on a pyrimidine compound.Suitable anions include chloride, bromide, iodide, sulfate, nitrate,phosphate, citrate, methanesulfonate, trifluoroacetate, acetate, malate,tosylate, tartrate, fumurate, glutamate, glucuronate, lactate,glutarate, and maleate. Likewise, a salt can also be formed between acation and a negatively charged group (e.g., carboxylate) on apyrimidine compound. Suitable cations include sodium ion, potassium ion,magnesium ion, calcium ion, and an ammonium cation such astetramethylammonium ion. The pyrimidine compounds also include thosesalts containing quaternary nitrogen atoms. Examples of prodrugs includeesters and other pharmaceutically acceptable derivatives, which, uponadministration to a subject, are capable of providing active pyrimidinecompounds. A solvate refers to a complex formed between an activepyrimidine compound and a pharmaceutically acceptable solvent. Examplesof pharmaceutically acceptable solvents include water, ethanol,isopropanol, ethyl acetate, acetic acid, and ethanol amine.

Also within the scope of this invention is a composition containing oneor more of the pyrimidine compounds described above for use in treatingan above-described disease, and the use of such a composition for themanufacture of a medicament for the just-mentioned treatment.

The details of one or more embodiments of the invention are set forth inthe description below. Other features, objects, and advantages of theinvention will be apparent from the description and from the claims.

DETAILED DESCRIPTION

Shown below are exemplary compounds, compounds 1-268, of this invention:

The pyrimidine compounds described above can be prepared by methods wellknown in the art. Examples 1-268 below provide detailed descriptions ofthe preparation of compounds 1-268 of this invention.

Scheme I shown below depicts a typical synthetic route for synthesizingcertain exemplary compounds. In this scheme, R₁, R₂, R₃, R₆, and R₇ areas defined in the Summary section above. Specifically, a pyrimidinecompound containing a halo group reacts with a compound containing twoprotected amino groups and an unprotected primary amino group to give acompound of formula (2), which is subsequently deprotected by removingthe amino-protecting group to give a compound of formula (3). Exemplaryamino-protecting groups include t-butoxycarbonyl, benzyloxycarbonyl,acetyl, phenylcarbonyl, and trialkylsilyl.

Compounds of formula (2) can be modified in various manners to affordother compounds of this invention. For example, as shown in Scheme IIbelow, when R₃ is halo, a compound of formula (2) reacts with aheterocyclic compound containing a ring nitrogen atom to give a compoundof formula (4), which is subsequently deprotected to give a compound offormula (5). As another example, when R₃ is also halo, a compound offormula (2) reacts with an alcohol to give a compound of formula (8),which is subsequently deprotected to give a compound of formual (9).

As shown in Scheme III below, when R₂ is CN, a compound of formula (2)can be first reduced to give a compound of formula (10), which containsan aldehyde group. The compound of formula (10) can then react with aprimary amine to give a compound of formula (11), which can besubsequently deprotected to form a compound of formula (12).

Compounds of formula (4) obtained above can be further modified invarious manners to give other compounds of this invention. For example,as shown in Scheme IV below, when Y is NH, a compound of formula (4)reacts with a compound containing a halide group, an aldehyde group, oran acyl chloride group to give a compound of formula (6), which issubsequently deprotected to give a compound of formula (7). As anotherexample, when Y is NH, a compound of formula (4) reacts with anα,β-unsaturated ester, followed by hydrolysis to give a compound offormula (13), which is subsequently deprotected to give a compound offormula (14). As another example, when Y is NH, a compound of formula(4) reacts with a vinyl phosphonate to give a compound of formula (15).The compound of formula (15) is then deprotected and hydrolyzed to givea compound of formula (16).

A pyrimidine compound thus synthesized can be purified by a method suchas column chromatography, high-pressure liquid chromatography, orrecrystallization.

Other pyrimidine compounds can be prepared using other suitable startingmaterials through the above synthetic routes and others known in theart. The methods described above may also additionally include steps,either before or after the steps described specifically herein, to addor remove suitable protecting groups in order to ultimately allowsynthesis of the pyrimidine compounds. In addition, various syntheticsteps may be performed in an alternate sequence or order to give thedesired compounds. Synthetic chemistry transformations and protectinggroup methodologies (protection and deprotection) useful in synthesizingapplicable pyrimidine compounds are known in the art and include, forexample, those described in R. Larock, Comprehensive OrganicTransformations, VCH Publishers (1989); T. W. Greene and P. G. M. Wuts,Protective Groups in Organic Synthesis, 2^(nd) Ed., John Wiley and Sons(1991); L. Fieser and M. Fieser, Fieser and Fieser's Reagents forOrganic Synthesis, John Wiley and Sons (1994); and L. Paquette, ed.,Encyclopedia of Reagents for Organic Synthesis, John Wiley and Sons(1995) and subsequent editions thereof.

The pyrimidine compounds mentioned herein may contain a non-aromaticdouble bond and one or more asymmetric centers. Thus, they can occur asracemates and racemic mixtures, single enantiomers, individualdiastereomers, diastereomeric mixtures, and cis- or trans-isomericforms. All such isomeric forms are contemplated.

Also within the scope of this invention is a pharmaceutical compositioncontaining an effective amount of at least one pyrimidine compounddescribed above and a pharmaceutical acceptable carrier. Further, thisinvention covers a method of administering an effective amount of one ormore of the pyrimidine compounds to a patient having a disease describedin the summary section above. This invention also covers a method ofadministering an effective amount of one or more of the pyrimidinecompounds for enhancing migration of bone marrow-derived cells to blood.“An effective amount” refers to the amount of an active pyrimidinecompound that is required to confer a therapeutic effect on the treatedsubject. Effective doses will vary, as recognized by those skilled inthe art, depending on the types of diseases treated, route ofadministration, excipient usage, and the possibility of co-usage withother therapeutic treatment.

To practice the method of the present invention, a composition havingone or more pyrimidine compounds can be administered parenterally,orally, nasally, rectally, topically, or buccally. The term “parenteral”as used herein refers to subcutaneous, intracutaneous, intravenous,intrmuscular, intraarticular, intraarterial, intrasynovial,intrasternal, intrathecal, intralesional, or intracranial injection, aswell as any suitable infusion technique.

A sterile injectable composition can be a solution or suspension in anon-toxic parenterally acceptable diluent or solvent, such as a solutionin 1,3-butanediol. Among the acceptable vehicles and solvents that canbe employed are mannitol, water, Ringer's solution, and isotonic sodiumchloride solution. In addition, fixed oils are conventionally employedas a solvent or suspending medium (e.g., synthetic mono- ordiglycerides). Fatty acid, such as oleic acid and its glyceridederivatives are useful in the preparation of injectables, as are naturalpharmaceutically acceptable oils, such as olive oil or castor oil,especially in their polyoxyethylated versions. These oil solutions orsuspensions can also contain a long chain alcohol diluent or dispersant,carboxymethyl cellulose, or similar dispersing agents. Other commonlyused surfactants such as Tweens or Spans or other similar emulsifyingagents or bioavailability enhancers which are commonly used in themanufacture of pharmaceutically acceptable solid, liquid, or otherdosage forms can also be used for the purpose of formulation.

A composition for oral administration can be any orally acceptabledosage form including capsules, tablets, emulsions and aqueoussuspensions, dispersions, and solutions. In the case of tablets,commonly used carriers include lactose and corn starch. Lubricatingagents, such as magnesium stearate, are also typically added. For oraladministration in a capsule form, useful diluents include lactose anddried corn starch. When aqueous suspensions or emulsions areadministered orally, the active ingredient can be suspended or dissolvedin an oily phase combined with emulsifying or suspending agents. Ifdesired, certain sweetening, flavoring, or coloring agents can be added.

A nasal aerosol or inhalation composition can be prepared according totechniques well known in the art of pharmaceutical formulation. Forexample, such a composition can be prepared as a solution in saline,employing benzyl alcohol or other suitable preservatives, absorptionpromoters to enhance bioavailability, fluorocarbons, and/or othersolubilizing or dispersing agents known in the art.

A composition having one or more active pyrimidine compounds can also beadministered in the form of suppositories for rectal administration.

The carrier in the pharmaceutical composition must be “acceptable” inthe sense that it is compatible with the active ingredient of thecomposition (and preferably, capable of stabilizing the activeingredient) and not deleterious to the subject to be treated. One ormore solubilizing agents can be utilized as pharmaceutical excipientsfor delivery of an active pyrimidine compound. Examples of othercarriers include colloidal silicon oxide, magnesium stearate, cellulose,sodium lauryl sulfate, and D&C Yellow # 10.

The pyrimidine compounds described above can be preliminarily screenedfor their efficacy in treating above-described diseases by an in vitroassay (See Examples 269 and 270 below) and then confirmed by animalexperiments and clinic trials. Other methods will also be apparent tothose of ordinary skill in the art.

The specific examples below are to be construed as merely illustrative,and not limitative of the remainder of the disclosure in any waywhatsoever. Without further elaboration, it is believed that one skilledin the art can, based on the description herein, utilize the presentinvention to its fullest extent. All publications cited herein arehereby incorporated by reference in their entirety.

EXAMPLE 1 Preparation of Compound 1

Hexamethyleneimine (0.673 g) was slowly added to a stirred solution of2,4-dichloropyrimidine (1 g) in THF (50 mL) at 0° C. The reactionmixture was stirred at 0° C. for 2 hours and the reaction was allowed towarm-up to room temperature overnight. The solution was thenconcentrated to give a residue, which was purified by chromatography onsilica gel (EtOAc/Hexane=1/5) to afford intermediate 1-I (1.234 g) in a86% yield.

A solution of Intermediate 1-I (0.46 g) and tris-(2-aminoethyl)-amine(1.6 g) in DMSO (2 mL) was heated at 120° C. for 8 hours by microwave.The solution was partitioned between CH₂Cl₂ and H₂O. The organic layerwas isolated and then concentrated. The residue was purified bychromatography on silica gel (21% NH₃(aq)/MeOH=1/3) to affordintermediate 1-II (0.454 g) in a 65% yield.

Cyclohexanone (488 mg) and NaBH(OAc)₃ (1320 mg) were added to a stirredsolution of intermediate 1-II (200 mg) in CH₂Cl₂ (50 mL) at roomtemperature over a short period of time. The resulting solution wasstirred at room temperature for 8 hours, and then quenched with asaturated aqueous NaHCO₃ solution. The aqueous layer was separated andextracted with CH₂Cl₂. The combined organic layers were subsequentlywashed with water, dried, filtered, and concentrated to give a cruderesidue, which was purified by chromatography on silica gel (21%NH₃(aq)/MeOH=1/10) to give compound 1 (217 mg) in a 72% yield.

CI-MS (M⁺+1): 486.4.

1 M hydrochloric acid (6 mL) and CH₂Cl₂ (4 ml) were added to thecompound 1 (217 mg). The mixture was stirred for 10 minutes at roomtemperature. After removing the supernatant, the solid was dried undervacuum to afford the hydrochloride salt of compound 1 (268 mg) in a 95%yield.

EXAMPLE 2 Preparation of Compound 2

Intermediate 2-I was obtained during the preparation of compound 1.

A solution of Intermediate 2-I (200 mg) and tris-(2-aminoethyl)amine (70mg) in DMSO (2 mL) was heated at 120° C. for 8 hours by microwave. Thesolution was partitioned between CH₂Cl₂ and H₂O. The organic layer wasisolated and then concentrated. The residue was purified bychromatography on silica gel (21% NH₃(aq)/MeOH=1/5) to affordintermediate 2-II (296 mg) in a 63% yield.

Cyclohexanone (234 mg) and NaBH(OAc)₃ (506 mg) were added to a stirredsolution of intermediate 2-II (296 mg) in CH₂Cl₂ (30 mL) at roomtemperature over a short period of time. The resulting solution wasstirred at room temperature for 8 hours, and then quenched with asaturated aqueous NaHCO₃ solution. The aqueous layer was separated andextracted with CH₂Cl₂. The combined organic layers were subsequentlywashed with water, dried, filtered, and concentrated to give a cruderesidue, which was purified by chromatography on silica gel (21%NH₃(aq)/MeOH=1/15) to give compound 2 (266 mg) in a 77% yield.

CI-MS (M⁺+1): 579.4.

1 M hydrochloric acid (6 mL) and CH₂Cl₂ (4 ml) were added to compound 2(266 mg). The mixture was stirred for 10 minutes at room temperature.After removing the supernatant, the solid was dried under vacuum toafford the hydrochloride salt of compound 2 (302 mg) in a 91% yield.

EXAMPLE 3 Preparation of Compound 3

Compound 3 was prepared in a manner similar to that used to preparecompound 1.

CI-MS (M⁺+1): 472.4.

EXAMPLE 4 Preparation of Compound 4

Compound 4 was prepared in a manner similar to that used to preparecompound 1.

CI-MS (M⁺+1): 514.4.

EXAMPLE 5 Preparation of Compound 5

A solution of tris-(2-aminoethyl)-amine (2.0 g) and Boc₂O (1.0 g) inCH₂Cl₂ (280 mL) was stirred at 25° C. for 15 hours and thenconcentrated. The resultant residue was purified by chromatography onsilica gel (EtOAc/MeOH=1/1) to afford intermediate 5-I (2.04 g) in a 43%yield.

1-Benzyl-4-piperidone (2.177 g) and NaBH(OAc)₃ (3.665 g) were added to astirred solution of intermediate 5-I (2.0 g) in MeOH (30 mL) at roomtemperature over a short period of time. The resulting solution wasstirred at room temperature for 8 hours, and then quenched with asaturated aqueous NaHCO₃ solution. The aqueous layer was separated andextracted with CH₂Cl₂. The combined organic layers were subsequentlywashed with water, dried, filtered, and concentrated to give a cruderesidue, which was purified by chromatography on silica gel(EtOAc/MeOH=9/1) to afford intermediate 5-II (2.488 g) in a 83% yield.

A solution of 20% TFA/CH₂Cl₂ (20 mL) was added to Intermediate 5-II (1.0g) in CH₂Cl₂ (10 mL). The reaction mixture was stirred for 5 hours atroom temperature and concentrated by removing the solvent. The residuewas purified by chromatography on silica gel (21% NH₃(aq)/MeOH=1/4) toafford intermediate 5-III (0.54 g) in a 88% yield.

Cyclohexanone (1,323 mg) and NaBH(OAc)₃ (3,220 mg) were added to astirred solution of intermediate 5-III (540 mg) in CH₂Cl₂ (30 mL) atroom temperature over a short period of time. The resulting solution wasstirred at room temperature for 8 hours and then quenched with asaturated aqueous NaHCO₃ solution. The aqueous layer was separated andextracted with CH₂Cl₂. The combined organic layers were subsequentlywashed with water, dried, filtered, and concentrated to give a cruderesidue, which was purified by chromatography on silica gel (21%NH₃(aq)/MeOH=1/10) to afford intermediate 5-IV (0.58 g) in a 71% yield.

A solution of intermediate 5-IV (580 mg), Boc₂O (863 mg) and Et₃N (485mg) in CH₂Cl₂ (150 ml) was stirred at 25° C. for 15 hours and thenconcentrated. The resultant residue was purified by chromatography onsilica gel (EtOAc/Hexane=1/5) to afford intermediate 5-V (865 mg) in a92% yield.

A mixture of intermediate 5-V (865 mg) and Pd/C (90 mg) in MeOH (20 ml)was stirred under H₂ (balloon) at 25° C. for 15 hours and then filteredthrough a celite column and concentrated. The resultant residue waspurified by chromatography on silica gel (EtOAc/MeOH=15/1) to affordintermediate 5-VI (681 mg) in a 89% yield.

Diisopropylethylamine (0.1 mL) was added to a solution of 5-VII (30 mg;obtained during preparation of compound 1) and intermediate 5-VI (100mg) in 1-pentanol (2 mL). The reaction mixture was stirred overnight at140° C. The solvent was then removed under vacuum and the resultantresidue was purified by chromatography on silica gel (EtOAc/Hexane=1/1)to afford intermediate 5-VIII (76 mg) in a 61% yield.

A solution of 20% TFA/CH₂Cl₂ (2 mL) was added to intermediate 5-VIII (76mg) in CH₂Cl₂ (1 mL). The reaction mixture was stirred for 5 hours atroom temperature and concentrated by removing the solvent. 1 Mhydrochloric acid (2 mL) and CH₂Cl₂ (1 mL) were added to the resultantresidue. The mixture was stirred for 10 minutes at room temperature.After removal of the supernatant, the solid was dried under vacuum toafford the hydrochloride salt of compound 5 (81 mg) in a 85% yield.

CI-MS (M⁺+1): 569.5.

EXAMPLE 6 Preparation of Compound 6

Compound 6 was prepared in a manner similar to that used to preparecompound 1.

CI-MS (M⁺+1): 572.5.

EXAMPLE 7 Preparation of Compound 7

Compound 7 was prepared in a manner similar to that used to preparecompound 1.

CI-MS (M⁺+1): 458.4.

EXAMPLE 8 Preparation of Compound 8

A solution of 4-cyanobenzylaldehyde (5 g) andN-cyclohexyl-1,3-propane-diamine (6 g) in CH₃OH (100 mL) was heated to60° C. for 6 hours. After cooling to room temperature, NaBH₄ (2.5 g) wasslowly added to the above solution. The mixture was stirred for another30 minutes. The mixture was then concentrated, quenched with NH₄Cl (aq),and extracted with CH₂Cl₂. The organic layers were combined, dried withanhydrous MgSO₄, and concentrated to give a residue. The residue waspurified by chromatography on silica gel (EtOAc/Et₃N=4/1) to affordIntermediate 8-I (7.2 g) in a 70% yield.

A solution of Intermediate 8-I (7.2 g) and Boc₂O (17.3 g) in CH₂Cl₂ (280ml) was stirred at 25° C. for 15 hours and then concentrated. Theresultant residue was purified by chromatography on silica gel(EtOAc/Hexane=1/1) to afford Intermediate 8-II as a yellow oil (10.6 g,yield: 85%).

A solution of Intermediate 8-II (4.7 g) and NiCl₂ (64 mg) in CH₃OH (100ml) was first stirred at 25° C. After cooling to 0° C., NaBH₄ (1.83 g)was slowly added and the mixture was stirred for another 15 hours. Thesolution was concentrated, quenched with NH₄Cl (aq), and extracted withCH₂Cl₂. The combined organic layer was washed with water, filtered,dried with anhydrous MgSO₄, and concentrated to give a residue. Theresidue was purified by chromatography on silica gel (21%NH₃(aq)/MeOH=1/19) to afford Intermediate 8-III (2.36 g) in a 50% yield.

Diisopropylethylamine (0.1 mL) was added to a solution of2-chloro-6-methyl-4-dimethylaminopyrimidine (110 mg) and Intermediate8-III (150 mg) in 1-pentanol (2 mL). The reaction mixture was stirredovernight at 150° C. The solvent was removed under vacuum and theresidue was purified by chromatography on silica gel (EtOAc/Hexane=1/1)to afford Intermediate 8-IV (88 mg) in a 47% yield.

A solution of 20% TFA/CH₂Cl₂ (2 mL) was added to Intermediate 8-IV (88mg) in CH₂Cl₂ (1 mL). The reaction mixture was stirred for 5 hours atroom temperature and concentrated by removing the solvent. 1 Mhydrochloric acid (2 mL) and CH₂Cl₂ (1 mL) were added to the resultantresidue. The mixture was stirred for 10 minutes at room temperature.After removal of the supernatant, the solid was dried under vacuum toafford the hydrochloride salt of compound 8 (60 mg) in an 80% yield.

CI-MS (M⁺+1): 411.3.

EXAMPLE 9 Preparation of Compound 9

A solution of N-cyclohexyl-1,3-propanediamine (2.6 g), Et₃N (3.8 mL) and2,4,6-trichloropyrimidine (1 g) in THF (50 mL) was stirred for overnightat 60° C. and concentrated by removing the solvent under vacuum. Theresidue was purified by chromatography on silica gel (21% NH₃(aq)/MeOH=5/95) to afford compound 9 (1.7 g) in a 75% yield.

CI-MS (M⁺+1): 423.3.

1 M hydrochloric acid (3 mL) and CH₂Cl₂ (2 ml) were added to compound 9(100 mg). The mixture was stirred for 10 minutes at room temperature.After removing the supernatant, the solid was dried under vacuum toafford the hydrochloride salt of compound 9 (130 mg) in a 97% yield.

EXAMPLE 10 Preparation of Compound 10

N-cyclohexyl-1,3-propanediamine (0.808 g) was slowly added to a stirredsolution of 2,4,6-trichloropyrimidine (1 g) in THF (50 mL) at roomtemperature. The reaction mixture was stirred at 0° C. for 2 hours andthe reaction was allowed to warm-up to room temperature overnight. Thesolution was then concentrated to give a residue, which was purified bychromatography on silica gel (EtOAc/Hexane=1/2) to afford Intermediate10-I (1.386 g) in a 60% yield.

A solution of Intermediate 10-I (500 mg) and Boc₂O (770 mg) in CH₂Cl₂(15 mL) was stirred at 25° C. overnight. The solution was thenconcentrated and the resultant residue was purified by chromatography onsilica gel (EtOAc/Hexane=1/9) to afford Intermediate 10-II (590 mg) inan 80% yield.

Diisopropylethylamine (0.25 mL) was added to a solution of Intermediate10-II (590 mg), Intermediate 8-III prepared in Example 8 (700 mg), andNaI (260 mg) in 1-pentanol (20 mL). The reaction mixture was stirred for24 hours at 120° C. and then concentrated by removing the solvent undervacuum. The resultant residue was dissolved in CH₂Cl₂, washed withwater, dried with anhydrous MgSO₄, and concentrated to give a residue.The residue was purified by chromatography on silica gel(EtOAc/Hexane=1/1) to afford Intermediate 10-III (865 mg) in a 70%yield.

A solution of 20% TFA/CH₂Cl₂ (3 mL) was added to a solution ofIntermediate 10-III (150 mg) in CH₂Cl₂ (2 mL). The reaction mixture wasstirred for 5 hours at room temperature and concentrated by removing thesolvent. 1 M hydrochloric acid (3 mL) and CH₂Cl₂ (2 mL) were added tothe residue. The mixture was stirred for another 10 minutes at roomtemperature. After removing the supernatant, the solid was dried undervacuum to afford the hydrochloride salt of compound 10 (107 mg) in an80% yield.

CI-MS (M⁺+1): 542.4.

EXAMPLE 11 Preparation of Compound 11

N-cyclohexyl-1,3-propanediamine (0.808 g) was slowly added to a stirredsolution of 2,4,6-trichloropyrimidine (1 g) in THF (50 mL) at roomtemperature. The reaction mixture was stirred at 0° C. for 2 hours andthe reaction was allowed to warm-up to room temperature overnight. Thesolution was then concentrated to give a residue, which was purified bychromatography on silica gel (EtOAc/Hexane=1/4) to afford intermediate11-I (0.349 g) in a 21% yield.

A solution of intermediate 11-I (349 mg) and Boc₂O (540 mg) in CH₂Cl₂(15 mL) was stirred at 25° C. overnight. The solution was thenconcentrated and the resultant residue was purified by chromatography onsilica gel (EtOAc/Hexane=1/10) to afford intermediate 11-II (400 mg) inan 86% yield (CI-MS (M⁺+1): 403.4).

Diisopropylethylamine (0.17 mL) was added to a solution of intermediate11-II (400 mg), intermediate 8-III (prepared in Example 8) (475 mg), andNaI (176 mg) in 1-pentanol (20 mL). The reaction mixture was stirred for24 hours at 120° C. and then concentrated by removing the solvent undervacuum. The resultant residue was dissolved in CH₂Cl₂, washed withwater, dried with anhydrous MgSO₄, and concentrated to give a residue.The residue was purified by chromatography on silica gel(EtOAc/Hexane=1/1) to afford Intermediate 11-III (427 mg) in a 51%yield.

A solution of 20% TFA/CH₂Cl₂ (4 mL) was added to a solution ofintermediate 11-III (200 mg) in CH₂Cl₂ (2 mL). The reaction mixture wasstirred for 5 hours at room temperature and concentrated by removing thesolvent. 1 M hydrochloric acid (5 mL) and CH₂Cl₂ (2 mL) were added tothe residue. The mixture was stirred for another 10 minutes at roomtemperature. After removing the supernatant, the solid was dried undervacuum to afford the hydrochloride salt of compound 11 (117 mg) in a 91%yield.

CI-MS (M⁺+1): 542.4.

EXAMPLE 12 Preparation of Compound 12

Compound 12 was prepared in a manner similar to that used to preparecompound 8.

CI-MS (M⁺+1): 422.

EXAMPLE 13 Preparation of Compound 13

Compound 13 was prepared in a manner similar to that used to preparecompound 10.

CI-MS (M⁺+1): 508.4.

EXAMPLE 14 Preparation of Compound 14

Compound 14 was prepared in a manner similar to that used to preparecompound 8.

CI-MS (M⁺+1): 387.

EXAMPLE 15 Preparation of Compound 15

Compound 15 was prepared in a manner similar to that used to preparecompound 8.

CI-MS (M⁺+1): 403.

EXAMPLE 16 Preparation of Compound 16

Compound 16 was prepared in a manner similar to that used to preparecompound 8.

CI-MS (M⁺+1): 354.3.

EXAMPLE 17 Preparation of Compound 17

Compound 17 was prepared in a manner similar to that used to preparecompound 11.

CI-MS (M⁺+1): 522.4.

EXAMPLE 18 Preparation of Compound 18

Compound 18 was prepared in a manner similar to that used to preparecompound 10.

CI-MS (M⁺+1): 522.4.

EXAMPLE 19 Preparation of Compound 19

Compound 19 was prepared in a manner similar to that used to preparecompound 11.

CI-MS (M⁺+1): 522.4.

EXAMPLE 20 Preparation of Compound 20

Intermediate 20-I was obtained as an intermediate during the preparationof compound 15.

NaH (110 mg) was added to a solution of the Intermediate 20-I (200 mg)and phenol (250 mg) in DMSO (3 mL). The reaction mixture was stirred at25° C. for 1 hour. The mixture was then heated at 120° C. for 8 hoursunder microwave, cooled to room temperature, and concentrated byremoving the solvent. The resultant residue was dissolved in CH₂Cl₂,washed with saturated aqueous NaHCO₃, dried with anhydrous MgSO₄, andconcentrated to give a residue. The residue was purified bychromatography on silica gel (EtOAc/Hexane=1/1) to afford Intermediate20-II (65 mg) in a 30% yield.

Intermediate 20-II (65 mg) in CH₂Cl₂ (1 mL) was added to a solution of20% TFA/CH₂Cl₂ (2 mL). The reaction mixture was stirred for 5 hours atroom temperature and concentrated by removing the solvent. 1 Mhydrochloric acid (2 mL) and CH₂Cl₂ (1 mL) were subsequently added tothe residue. The mixture was stirred for 10 minutes at room temperature.After removing the supernatant, the solid was dried under vacuum toafford the hydrochloride salt of compound 20 (45 mg) in an 80% yield.

CI-MS (M⁺+1): 461.3.

EXAMPLE 21 Preparation of Compound 21

Intermediate 21-I was obtained as an intermediate during the preparationof compound 20.

A solution of 21-I (2 g) and piperazine (10 g) in 1-pentanol (3 mL) wasstirred for 4 hours at 120° C. and concentrated by removing the solventunder vacuum. The resultant mixture was dissolved in CHCl₃, washed withwater, dried with anhydrous MgSO₄, and concentrated to give a residue.The residue was purified by chromatography on silica gel (21% NH₃(aq)/MeOH=1/99) to afford Intermediate 21-II (1.5 g) in a 60% yield.

A solution of 20% TFA/CH₂Cl₂ (3 mL) was added to a solution ofIntermediate 20-II (130 mg) in CH₂Cl₂ (2 mL). The reaction mixture wasstirred for 5 hours at room temperature and then concentrated byremoving the solvent. 1 M hydrochloric acid (3 mL) and CH₂Cl₂ (2 ml)were added to the resultant residue. The mixture was stirred for another10 minutes at room temperature. After removing the supernatant, thesolid was dried under vacuum to afford the hydrochloride salt ofcompound 21 (90 mg) in an 80% yield.

CI-MS (M⁺+1): 453.3.

EXAMPLE 22 Preparation of Compound 22

Compound 22 was prepared in a manner similar to that used to preparecompound 21.

CI-MS (M⁺+1): 497.4

EXAMPLE 23 Preparation of Compound 23

Compound 23 was prepared in a manner similar to that used to preparecompound 21.

CI-MS (M⁺+1): 467.4.

EXAMPLE 24 Preparation of Compound 24

Compound 24 was prepared in a manner similar to that used to preparecompound 10.

CI-MS (M⁺+1): 522.4.

EXAMPLE 25 Preparation of Compound 25

Compound 25 was prepared in a manner similar to that used to preparecompound 11.

CI-MS (M⁺+1): 466.4.

EXAMPLE 26 Preparation of Compound 26

Compound 26 was prepared in a manner similar to that used to preparecompound 8.

CI-MS (M⁺+1): 394.3.

EXAMPLE 27 Preparation of Compound 27

Compound 27 was prepared in a manner similar to that used to preparecompound 21.

CI-MS (M⁺+1): 454.3.

EXAMPLE 28 Preparation of Compound 28

Compound 28 was prepared in a manner similar to that used to preparecompound 21.

CI-MS (M⁺+1): 452.3.

EXAMPLE 29 Preparation of Compound 29

Intermediate 29-I was obtained as an intermediate during the preparationof compound 26.

1M DIBAL/ether (8.35 mL) was added to a stirred solution of Intermediate29-I (1.24 g) in dry toluene (100 mL) at −70˜−78° C. under N₂ (g). Thereaction mixture was stirred for 2 hours at this temperature. 5% HCl(aq) (9 mL) was then added to the solution at −60˜−70° C. and themixture was stirred for another 0.5 hour after the reaction temperaturewas increased to 25° C. To the solution was added CH₂Cl₂ (100 mL) andH₂O. The aqueous layer was extracted with CH₂Cl₂ twice. The organiclayers were combined, dried with anhydrous MgSO₄, and concentrated byremoving the solvent under vacuum. The resultant residue was purified bychromatography on silica gel (EtOAc/Hexane=1/2) to afford Intermediate29-II (620 mg) in a 50% yield.

A solution of tryptamine (99 mg) and Intermediate 29-II (170 mg) inCH₃OH (6 mL) was heated at 60° C. for 6 hours. After cooling to roomtemperature, NaBH₄ (20 mg) was slowly added to the solution and themixture was stirred for another 30 minutes. The mixture wasconcentrated, quenched with NH₄Cl (aq), extracted with CH₂Cl₂. Theorganic layer was dried with anhydrous MgSO₄ and concentrated to give aresidue. The residue was purified by chromatography on silica gel(EtOAc/MeOH=9/1) to afford Intermediate 29-III (150 mg) in a 70% yield.

Intermediate 29-III (150 mg) in CH₂Cl₂ (2 mL) was added to a solution of20% TFA/CH₂Cl₂ (3 mL). The reaction mixture was stirred for 5 hours atroom temperature and concentrated by removing the solvent. 1 Mhydrochloric acid (3 mL) and CH₂Cl₂ (2 mL) were added to the residueobtained above. The mixture was stirred for another 10 minutes at roomtemperature. After removal of the supernatant, the solid was dried undervacuum to afford the hydrochloride salt of compound 29 (92 mg) in a 70%yield.

CI-MS (M⁺+1): 541.4.

EXAMPLE 30 Preparation of Compound 30

Compound 30 was prepared in a manner similar to that used to preparecompound 29.

CI-MS (M⁺+1): 528.3.

EXAMPLE 31 Preparation of Compound 31

Compound 31 was prepared in a manner similar to that used to preparecompound 21.

CI-MS (M⁺+1): 481.4.

EXAMPLE 32 Preparation of Compound 32

Compound 32 was prepared in a manner similar to that used to preparecompound 21.

CI-MS (M⁺+1): 547.4.

EXAMPLE 33 Preparation of Compound 33

Intermediate 33-I was obtained as an intermediate during the preparationof compound 21.

Diisopropylethylamine (0.1 mL) and cyclohexanecarbonyl chloride (55 mg)were added to a solution of Intermediate 33-I (200 mg) in CH₂Cl₂ (10mL). The reaction mixture was stirred overnight at room temperature andthen concentrated by removing the solvent. The resultant mixture wasdissolved in CHCl₃, washed with water, dried with anhydrous MgSO₄, andconcentrated to give a residue. The residue was purified bychromatography on silica gel (EtOAc/Hexane=1/1) to afford Intermediate33-II (140 mg) in a 60% yield.

A solution of 20% TFA/CH₂Cl₂ (3 mL) was added to a solution ofIntermediate 33-II (140 mg) in CH₂Cl₂ (2 mL). The reaction mixture wasstirred for 5 hours at room temperature and concentrated by removing thesolvent. 1 M hydrochloric acid (3 mL) and CH₂Cl₂ (2 mL) were added tothe residue. The resultant mixture was stirred for another 10 minutes atroom temperature. After removal of the supernatant, the solid was driedunder vacuum to afford the hydrochloride salt of compound 33 (100 mg) inan 80% yield.

CI-MS (M⁺+1): 563.4.

EXAMPLE 34 Preparation of Compound 34

Intermediate 34-I was obtained as an intermediate during the preparationof compound 21. Intermediate 34-I (166 mg) was first dissolved in CH₃CN(10 mL). 2-chloromethylbenzimidazole (42 mg) and K₂CO₃ (79 mg) were thenadded to the above solution. After the mixture was stirred for 48 hoursat room temperature, it was filtered and concentrated. The resultantresidue was purified by chromatography on silica gel (EtOAc/MeOH=10/1)to afford Intermediate 34-II (70 mg) in a 35% yield.

A solution of 20% TFA/CH₂Cl₂ (2 mL) was added to a solution ofIntermediate 34-II (70 mg) in CH₂Cl₂ (1 mL). The reaction mixture wasstirred for 5 hours at room temperature and concentrated by removing thesolvent. 1 M hydrochloric acid (2 mL) and CH₂Cl₂ (1 mL) were added tothe resultant residue. The mixture was stirred for another 10 minutes atroom temperature. After removal of the supernatant, the solid was driedunder vacuum to afford the hydrochloride salt of compound 34 (50 mg) inan 80% yield.

CI-MS (M⁺+1): 583.4.

EXAMPLE 35 Preparation of Compound 35

Intermediate 35-I was obtained as an intermediate during the preparationof compound 21.

NaBH(OAc)₃ (215 mg) was added to a solution of Intermediate 35-I (166mg) in MeOH (10 mL) and cyclohexanecarbaldehyde (57 mg). A few drops ofacetic acid was then added. The reaction mixture was stirred for 48hours at room temperature and concentrated by removing the solventthrough distillation. The resultant mixture was dissolved in CH₂Cl₂ andquenched with saturated aqueous NaHCO₃. The aqueous layer was separatedand extracted with CH₂Cl₂. The combined organic layers were subsequentlywashed with water, dried with anhydrous MgSO₄, filtered, andconcentrated to give a residue. The residue was purified bychromatography on silica gel (EtOAc/Hexane=2/1) to give Intermediate35-II (120 mg) in a 65% yield.

A solution of 20% TFA/CH₂Cl₂ (3 mL) was added to a solution ofIntermediate 35-II (120 mg) in CH₂Cl₂ (2 mL). The reaction mixture wasstirred for 5 hours at room temperature and concentrated by removing thesolvent. 1 M hydrochloric acid (3 mL) and CH₂Cl₂ (2 mL) were added tothe residue. The mixture was stirred for another 10 minutes at roomtemperature. After removal of the supernatant, the solid was dried undervacuum to afford the hydrochloride salt of compound 35 (85 mg) in an 80%yield.

CI-MS (M⁺+1): 549.4.

EXAMPLE 36 Preparation of Compound 36

Compound 36 was prepared in a manner similar to that used to preparecompound 35.

CI-MS (M⁺+1): 543.4.

EXAMPLE 37 Preparation of Compound 37

Compound 37 was prepared in a manner similar to that used to preparecompound 21.

CI-MS (M⁺+1): 563.4.

EXAMPLE 38 Preparation of Compound 38

Compound 38 was prepared in a manner similar to that used to preparecompound 21.

CI-MS (M⁺+1): 564.4.

EXAMPLE 39 Preparation of Compound 39

Compound 39 was prepared in a manner similar to that used to preparecompound 21.

CI-MS (M⁺+1): 566.4.

EXAMPLE 40 Preparation of Compound 40

Compound 40 was prepared in a manner similar to that used to preparecompound 21.

CI-MS (M⁺+1): 587.4.

EXAMPLE 41 Preparation of Compound 41

Compound 41 was prepared in a manner similar to that used to preparecompound 33.

CI-MS (M⁺+1): 523.4.

EXAMPLE 42 Preparation of Compound 42

Compound 42 was prepared in a manner similar to that used to preparecompound 33.

CI-MS (M⁺+1): 557.4.

EXAMPLE 43 Preparation of Compound 43

Intermediate 43-I was prepared in a 45% yield in a manner similar tothat described in the first paragraph of Example 10 using benzylamine(645 mg) as a starting material.

Diisopropylethylamine (0.27 mL) was added to a solution of Intermediate43-I (625 mg), Intermediate 8-III prepared in Example 8 (741 mg), andNaI (275 mg) in 1-pentanol (20 mL). The reaction mixture was stirred at120° C. for 24 hours and concentrated by removing the solvent undervacuum. The resultant mixture was dissolved in CH₂Cl₂, washed withwater, dried with anhydrous MgSO₄, and concentrated to give a residue.The residue was purified by chromatography on silica gel(EtOAc/Hexane=1/3) to afford Intermediate 43-II (1,100 mg) in a 65%yield.

A solution of Intermediate 43-II (200 mg) and N-methylpiperazine (2,000mg) in 1-pentanol (1 mL) was stirred at 120° C. for 4 hours. The solventwas then removed under vacuum. The resultant mixture was dissolved inCHCl₃, washed with water, dried with anhydrous MgSO₄, and concentratedto give a residue. The residue was purified by chromatography on silicagel (EtOAc/MeOH=20/1) to afford Intermediate 43-III (215 mg) in a 70%yield.

A solution of 20% TFA/CH₂Cl₂ (4 mL) was added to a solution ofIntermediate 43-III (215 mg) in CH₂Cl₂ (2 mL). The reaction mixture wasstirred for 5 hours at room temperature and concentrated by removing thesolvent. 1 M hydrochloric acid (4 mL) and CH₂Cl₂ (2 mL) were added tothe residue. The mixture was stirred for another 10 minutes at roomtemperature. After removal of the supernatant, the solid was dried undervacuum to afford the hydrochloride salt of compound 43 (150 mg) in an80% yield.

CI-MS (M⁺+1): 557.4.

EXAMPLE 44 Preparation of Compound 44

1-Methylpiperazine (0.76 g) and Et₃N (0.8 mL) were added to a solutionof 2,4-dichloro-6-methylpyrimidine (1 g) in EtOH (60 mL). The reactionsolution was stirred at 0° C. for 1 hour and then was allowed to warm-upto room temperature within 3 hours. The solution was then concentratedto give a residue, which was purified by chromatography on silica gel(EtOAc/MeOH=6/1) to afford Intermediate 44-I (0.76 g) in a 55% yield.

Diisopropylethylamine (0.25 mL) was added to a solution of Intermeidate44-I (300 mg), Intermediate 8-III prepared in Example 8 (689 mg), andNaI (260 mg) in 1-pentanol (20 mL). The reaction mixture was stirred for24 hours at 120° C. and concentrated by removing the solvent undervacuum. The resultant mixture was dissolved in CH₂Cl₂, washed withwater, dried with anhydrous MgSO₄, and concentrated to give a residue.The residue was purified by chromatography on silica gel(EtOAc/MeOH=5/1) to afford Intermediate 44-II (530 mg) in a 60% yield.

A solution of 20% TFA/CH₂Cl₂ (3 mL) was added to a solution of compoundIntermediate 44-II (150 mg) in CH₂Cl₂ (2 mL). The reaction mixture wasstirred for 5 hours at room temperature and concentrated by removing thesolvent. 1 M hydrochloric acid (3 mL) and CH₂Cl₂ (2 mL) were added tothe residue. The mixture was stirred for another 10 minutes at roomtemperature. After removal of the supernatant, the solid was dried undervacuum to afford the hydrochloride salt of compound 44 (100 mg) in an80% yield.

CI-MS (M⁺+1): 466.4.

EXAMPLE 45 Preparation of Compound 45

Compound 45 was prepared in a manner similar to that used to preparecompound 43.

CI-MS (M⁺+1): 493.3.

EXAMPLE 46 Preparation of Compound 46

Compound 46 was prepared in a manner similar to that used to preparecompound 43.

CI-MS (M⁺+1): 587.4.

EXAMPLE 47 Preparation of Compound 47

Compound 47 was prepared in a manner similar to that used to preparecompound 21.

CI-MS (M⁺+1): 549.4.

EXAMPLE 48 Preparation of Compound 48

Compound 48 was prepared in a manner similar to that used to preparecompound 21.

CI-MS (M⁺+1): 524.4.

EXAMPLE 49 Preparation of Compound 49

Compound 49 was prepared in a manner similar to that used to preparecompound 21.

CI-MS (M⁺+1): 521.4.

EXAMPLE 50 Preparation of Compound 50

Compound 50 was prepared in a manner similar to that used to preparecompound 43.

CI-MS (M⁺+1): 550.4.

EXAMPLE 51 Preparation of Compound 51

Compound 51 was prepared in a manner similar to that used to preparecompound 43.

CI-MS (M⁺+1): 580.4.

EXAMPLE 52 Preparation of Compound 52

Compound 52 was prepared in a manner similar to that used to preparecompound 43.

CI-MS (M⁺+1): 567.4.

EXAMPLE 53 Preparation of Compound 53

Compound 53 was prepared in a manner similar to that used to preparecompound 44.

CI-MS (M⁺+1): 542.4.

EXAMPLE 54 Preparation of Compound 54

Compound 54 was prepared in a manner similar to that used to preparecompound 44.

CI-MS (M⁺+1): 548.4.

EXAMPLE 55 Preparation of Compound 55

Compound 55 was prepared in a manner similar to that used to preparecompound 44.

CI-MS (M⁺+1): 520.4.

EXAMPLE 56 Preparation of Compound 56

Compound 56 was prepared in a manner similar to that used to preparecompound 44.

CI-MS (M⁺+1): 562.5.

EXAMPLE 57 Preparation of Compound 57

Compound 57 was prepared in a manner similar to that used to preparecompound 44.

CI-MS (M⁺+1): 508.4.

EXAMPLE 58 Preparation of Compound 58

Compound 58 was prepared in a manner similar to that used to preparecompound 44.

CI-MS (M⁺+1): 496.4.

EXAMPLE 59 Preparation of Compound 59

Compound 59 was prepared in a manner similar to that used to preparecompound 44.

CI-MS (M⁺+1): 534.4.

EXAMPLE 60 Preparation of Compound 60

Compound 60 was prepared in a manner similar to that used to preparecompound 44.

CI-MS (M⁺+1): 556.4.

EXAMPLE 61 Preparation of Compound 61

Compound 61 was prepared in a manner similar to that used to preparecompound 44.

CI-MS (M⁺+1): 618.4.

EXAMPLE 62 Preparation of Compound 62

1-(Diphenylmethyl)piperazine (3.78 g) and Et₃N (2.3 mL) were added to asolution of 2,4-dichloro-6-methylpyrimidine (2.43 g) in EtOH (200 mL).The reaction mixture was stirred at 0° C. for 1 hour and then wasallowed to warm-up to room temperature within 15 hours. The solution wasthen concentrated to give a residue, which was purified bychromatography on silica gel (EtOAc/Hexane=1/5) to afford intermediate62-I (4.0 g) in a 70% yield.

Diisopropylethylamine (4.09 g) was added to a solution of intermediate92-I (4.0 g), intermediate 8-III prepared in Example 8 (5.29 g), and NaI(2.38 g) in 1-pentanol (10 mL). The reaction mixture was stirred for 15hours at 140° C. and concentrated by removing the solvent under vacuum.The resultant mixture was dissolved in CH₂Cl₂, washed with water, driedwith anhydrous MgSO₄, and concentrated to give a residue. The residuewas purified by chromatography on silica gel (EtOAc/Hexane=1/1) toafford intermediate 62-II (2.7 g) in a 31% yield.

A solution of intermediate 62-II (2.7 g) and Pd/C (2.0 g) in isopropanol(30 ml) was stirred under H₂ (balloon) at 60° C. for 3 hours and thenfiltered through a celite column and concentrated. The resultant residuewas purified by chromatography on silica gel (EtOAc/MeOH=10/1) to affordintermediate 62-III (1.1 g) in a 50% yield.

Intermediate 62-III (200 mg) was first dissolved in CH₃CN (10 mL).2-chloromethylbenzimidazole (51 mg) and K₂CO₃ (86 mg) were then added tothe above solution. After the mixture was stirred for 48 hours at roomtemperature, it was filtered and concentrated. The resultant residue waspurified by chromatography on silica gel (EtOAc/MeOH=10/1) to affordintermediate 62-IV (100 mg) in a 42% yield.

A solution of 20% TFA/CH₂Cl₂ (3 mL) was added to a solution ofintermediate 62-IV (100 mg) in CH₂Cl₂ (2 mL). The reaction mixture wasstirred for 5 hours at room temperature and concentrated by removing thesolvent. 1 M hydrochloric acid (3 mL) and CH₂Cl₂ (2 mL) were added tothe residue. The mixture was stirred for another 10 minutes at roomtemperature. After removal of the supernatant, the solid was dried undervacuum to afford the hydrochloride salt of compound 62 (100 mg) in a 84%yield.

CI-MS (M⁺+1): 582.4.

EXAMPLE 63 Preparation of Compound 63

Compound 63 was prepared in a manner similar to that used to preparecompound 44.

CI-MS (M⁺+1): 572.4.

EXAMPLE 64 Preparation of Compound 64

Compound 64 was prepared in a manner similar to that used to preparecompound 44.

CI-MS (M⁺+1): 576.4.

EXAMPLE 65 Preparation of Compound 65

Compound 65 was prepared in a manner similar to that used to preparecompound 21.

CI-MS (M⁺+1): 525.4.

EXAMPLE 66 Preparation of Compound 66

Compound 66 was prepared in a manner similar to that used to preparecompound 21.

CI-MS (M⁺+1): 535.4.

EXAMPLE 67 Preparation of Compound 67

Compound 67 was prepared in a manner similar to that used to preparecompound 21.

CI-MS (M⁺+1): 569.4.

EXAMPLE 68 Preparation of Compound 68

Compound 68 was prepared in a manner similar to that used to preparecompound 21.

CI-MS (M⁺+1): 525.4.

EXAMPLE 69 Preparation of Compound 69

Compound 69 was prepared in a manner similar to that used to preparecompound 21.

CI-MS (M⁺+1): 547.3.

EXAMPLE 70 Preparation of Compound 70

Intermediate 70-I was obtained during the preparation of compound 21.

A solution of intermediate 70-I (150 mg), 3-phenoxyphenyl isocyanate (48mg) and Et₃N (46 mg) in CH₂Cl₂ (15 ml) was stirred at 25° C. overnightand then concentrated. The resultant residue was purified bychromatography on silica gel (EtOAc/MeOH=20/1) to afford Intermediate70-II (163 mg) in a 82% yield.

A solution of 20% TFA/CH₂Cl₂ (3 mL) was added to a solution ofintermediate 70-II (163 mg) in CH₂Cl₂ (2 mL). The reaction mixture wasstirred for 5 hours at room temperature and concentrated by removing thesolvent. 1 M hydrochloric acid (3 mL) and CH₂Cl₂ (2 mL) were added tothe residue. The mixture was stirred for another 10 minutes at roomtemperature. After removal of the supernatant, the solid was dried undervacuum to afford the hydrochloride salt of compound 70 (164 mg) in a 86%yield.

CI-MS (M⁺+1): 664.4.

EXAMPLE 71 Preparation of Compound 71

Compound 71 was prepared in a manner similar to that used to preparecompound 21.

CI-MS (M⁺+1): 597.4.

EXAMPLE 72 Preparation of Compound 72

Compound 72 was prepared in a manner similar to that used to preparecompound 21.

CI-MS (M⁺+1): 468.3.

EXAMPLE 73 Preparation of Compound 73

Compound 73 was prepared in a manner similar to that used to preparecompound 21.

CI-MS (M⁺+1): 530.4.

EXAMPLE 74 Preparation of Compound 74

Compound 74 was prepared in a manner similar to that used to preparecompound 35.

CI-MS (M⁺+1): 523.4.

EXAMPLE 75 Preparation of Compound 75

Compound 75 was prepared in a manner similar to that used to preparecompound 35.

CI-MS (M⁺+1): 537.4.

EXAMPLE 76 Preparation of Compound 76

Compound 76 was prepared in a manner similar to that used to preparecompound 44.

CI-MS (M⁺+1): 566.4.

EXAMPLE 77 Preparation of Compound 77

Compound 77 was prepared in a manner similar to that used to preparecompound 44.

CI-MS (M⁺+1): 386.

EXAMPLE 78 Preparation of Compound 78

Compound 78 was prepared in a manner similar to that used to preparecompound 44.

CI-MS (M⁺+1): 565.4.

EXAMPLE 79 Preparation of Compound 79

Compound 79 was prepared in a manner similar to that used to preparecompound 70.

CI-MS (M⁺+1): 640.4.

EXAMPLE 80 Preparation of Compound 80

Compound 80 was prepared in a manner similar to that used to preparecompound 33.

CI-MS (M⁺+1): 537.4.

EXAMPLE 81 Preparation of Compound 81

Compound 81 was prepared in a manner similar to that used to preparecompound 33.

CI-MS (M⁺+1): 537.4.

EXAMPLE 82 Preparation of Compound 82

Compound 82 was prepared in a manner similar to that used to preparecompound 33.

CI-MS (M⁺+1): 521.4.

EXAMPLE 83 Preparation of Compound 83

Compound 83 was prepared in a manner similar to that used to preparecompound 33.

CI-MS (M⁺+1): 586.4.

EXAMPLE 84 Preparation of Compound 84

Compound 84 was prepared in a manner similar to that used to preparecompound 44.

CI-MS (M⁺+1): 524.4.

EXAMPLE 85 Preparation of Compound 85

Compound 85 was prepared in a manner similar to that used to preparecompound 44.

CI-MS (M⁺+1): 546.3.

EXAMPLE 86 Preparation of Compound 86

Compound 86 was prepared in a manner similar to that used to preparecompound 70.

CI-MS (M⁺+1): 552.4.

EXAMPLE 87 Preparation of Compound 87

Compound 87 was prepared in a manner similar to that used to preparecompound 35.

CI-MS (M⁺+1): 523.4.

EXAMPLE 88 Preparation of Compound 88

Compound 88 was prepared in a manner similar to that used to preparecompound 35.

CI-MS (M⁺+1): 509.4.

EXAMPLE 89 Preparation of Compound 89

Compound 89 was prepared in a manner similar to that used to preparecompound 43.

CI-MS (M⁺+1): 475.3.

EXAMPLE 90 Preparation of Compound 90

Compound 90 was prepared in a manner similar to that used to preparecompound 44.

CI-MS (M⁺+1): 453.4.

EXAMPLE 91 Preparation of Compound 91

Compound 91 was prepared in a manner similar to that used to preparecompound 44.

CI-MS (M⁺+1): 494.4.

EXAMPLE 92 Preparation of Compound 92

Compound 92 was prepared in a manner similar to that used to preparecompound 34.

CI-MS (M⁺+1): 601.4.

EXAMPLE 93 Preparation of Compound 93

Compound 93 was prepared in a manner similar to that used to preparecompound 34.

CI-MS (M⁺+1): 535.4.

EXAMPLE 94 Preparation of Compound 94

Compound 94 was prepared in a manner similar to that used to preparecompound 33.

CI-MS (M⁺+1): 577.4.

EXAMPLE 95 Preparation of Compound 95

Intermediate 95-I was obtained during the preparation of compound 21.

A solution of intermediate 95-I (150 mg), trifluoroacetic anhydride (240mg), and Et₃N (230 mg) in CH₂Cl₂ (10 ml) was stirred at 25° C. overnightand then concentrated. The resultant residue was purified bychromatography on silica gel (EtOAc/MeOH=20/1) to afford Intermediate95-II (148 mg) in a 76% yield.

A solution of 20% TFA/CH₂Cl₂ (3 mL) was added to a solution of compound95-II (148 mg) in CH₂Cl₂ (2 mL). The reaction mixture was stirred for 5hours at room temperature and concentrated by removing the solvent. 1 Mhydrochloric acid (3 mL) and CH₂Cl₂ (2 mL) were added to the residue.The mixture was stirred for another 10 minutes at room temperature.After removal of the supernatant, the solid was dried under vacuum toafford the hydrochloride salt of compound 95 (127 mg) in a 92% yield.

CI-MS (M⁺+1): 645.3.

EXAMPLE 96 Preparation of Compound 96

Compound 96 was prepared in a manner similar to that used to preparecompound 43.

CI-MS (M⁺+1): 515.4.

EXAMPLE 97 Preparation of Compound 97

Compound 97 was prepared in a manner similar to that used to preparecompound 43.

CI-MS (M⁺+1): 519.3.

EXAMPLE 98 Preparation of Compound 98

Compound 98 was prepared in a manner similar to that used to preparecompound 43.

CI-MS (M⁺+1): 511.3.

EXAMPLE 99 Preparation of Compound 99

Compound 99 was prepared in a manner similar to that used to preparecompound 43.

CI-MS (M⁺+1): 459.3.

EXAMPLE 100 Preparation of Compound 100

Compound 100 was prepared in a manner similar to that used to preparecompound 43.

CI-MS (M⁺+1): 638.5.

EXAMPLE 101 Preparation of Compound 101

Compound 101 was prepared in a manner similar to that used to preparecompound 43.

CI-MS (M⁺+1): 605.5.

EXAMPLE 102 Preparation of Compound 102

Compound 102 was prepared in a manner similar to that used to preparecompound 43.

CI-MS (M⁺+1): 553.4.

EXAMPLE 103 Preparation of Compound 103

Compound 103 was prepared in a manner similar to that used to preparecompound 44.

CI-MS (M⁺+1): 549.4.

EXAMPLE 104 Preparation of Compound 104

Compound 104 was prepared in a manner similar to that used to preparecompound 44.

CI-MS (M⁺+1): 523.4.

EXAMPLE 105 Preparation of Compound 105

Compound 105 was prepared in a manner similar to that used to preparecompound 44.

CI-MS (M⁺+1): 524.4.

EXAMPLE 106 Preparation of Compound 106

Compound 106 was prepared in a manner similar to that used to preparecompound 33.

CI-MS (M⁺+1): 580.4.

EXAMPLE 107 Preparation of Compound 107

Compound 107 was prepared in a manner similar to that used to preparecompound 44.

CI-MS (M⁺+1): 580.4.

EXAMPLE 108 Preparation of Compound 108

Compound 108 was prepared in a manner similar to that used to preparecompound 44.

CI-MS (M⁺+1): 563.4.

EXAMPLE 109 Preparation of Compound 109

Intermediate 109-I was obtained during the preparation of compound 21.

A solution of intermediate 109-I (100 mg), isopropylsulfonyl chloride(35 mg), and Et₃N (30 mg) in CH₂Cl₂ (10 ml) was stirred at 25° C. forovernight and then concentrated. The resultant residue was purified bychromatography on silica gel (EtOAc/MeOH=20/1) to afford intermediate109-II (100 mg) in a 86% yield.

A solution of 20% TFA/CH₂Cl₂ (3 mL) was added to a solution of compound109-II (100 mg) in CH₂Cl₂ (2 mL). The reaction mixture was stirred for 5hours at room temperature and concentrated by removing the solvent. 1 Mhydrochloric acid (3 mL) and CH₂Cl₂ (2 mL) were added to the residue.The mixture was stirred for another 10 minutes at room temperature.After removal of the supernatant, the solid was dried under vacuum toafford the hydrochloride salt of compound 109 (80 mg) in a 87% yield.

CI-MS (M⁺+1): 559.3.

EXAMPLE 110 Preparation of Compound 110

Compound 110 was prepared in a manner similar to that used to preparecompound 70.

CI-MS (M⁺+1): 602.4.

EXAMPLE 111 Preparation of Compound 111

Compound 111 was prepared in a manner similar to that used to preparecompound 70.

CI-MS (M⁺+1): 582.4.

EXAMPLE 112 Preparation of Compound 112

Compound 112 was prepared in a manner similar to that used to preparecompound 44.

CI-MS (M⁺+1): 537.4.

EXAMPLE 113 Preparation of Compound 113

Compound 113 was prepared in a manner similar to that used to preparecompound 33.

CI-MS (M⁺+1): 563.4.

EXAMPLE 114 Preparation of Compound 114

4-(tert-butoxycarbonylamino-methyl)benzoic acid (1,000 mg) was dissolvedin CH₂Cl₂ (50 mL). N-cyclohexyl-1,3-propanediamine (745 mg),1-hydroxybenzo-triazole hydrate (HOBt, 645 mg), 4-methylmorpholine(1,607 mg), and 1-(3-(dimethylamino)propyl)-3-ethylcarbodiimidehydrochloride (EDC, 740 mg) were added to the above solutionsequentially. The reaction mixture was stirred at 25° C. for 18 hoursand then was partitioned between water (150 mL) and EtOAc (2×150 mL).The combined organic layers were dried over Na₂SO₄ and wereconcentrated. The resultant residue was purified by chromatography onsilica gel (EtOAc/MeOH=20/1) to afford intermediate 114-I (1,000 mg) ina 65% yield.

A solution of HCl in ether (1.0 M, 20 mL) was added to a solution of114-I (1,000 mg) in MeOH (10 mL) at 25° C. The mixture was stirred for12 hours at room temperature. After removal of the supernatant, thesolid was dried under vacuum to afford the hydrochloride salt ofintermediate 114-II (743 mg) in a 73% yield.

Diisopropylethylamine (134 mg) was added to a solution of intermediate114-II (250 mg), intermediate 90-I prepared in Example 90 (184 mg), andNaI (22 mg) in 1-pentanol (5 mL). The reaction mixture was stirred for15 hours at 140° C. and concentrated by removing the solvent undervacuum. The resultant mixture was dissolved in CH₂Cl₂, washed withwater, dried with anhydrous MgSO₄, and concentrated to give a residue.The residue was purified by chromatography on silica gel (21%NH₃(aq)/MeOH=1/15) to give compound 114 (213 mg) in a 53% yield.

CI-MS (M⁺+1): 467.3.

EXAMPLE 115 Preparation of Compound 115

Compound 115 was prepared in a manner similar to that used to preparecompound 44.

CI-MS (M⁺+1): 536.4.

EXAMPLE 116 Preparation of Compound 116

Intermediate 116-I was obtained during the preparation of compound 62.

Et₃N (0.18 mL) and cyclopropylcarbonyl chloride (96 mg) were added to asolution of intermediate 116-I (200 mg) in CH₂Cl₂ (10 mL). The reactionmixture was stirred overnight at room temperature and then concentratedby removing the solvent. The resultant mixture was dissolved in CHCl₃,washed with water, dried with anhydrous MgSO₄, and concentrated to givea residue. The residue was purified by chromatography on silica gel(EtOAc/Hexane=1/1) to afford intermediate 116-II (126 mg) in a 57%yield.

A solution of 20% TFA/CH₂Cl₂ (3 mL) was added to a solution ofintermediate 116-II (126 mg) in CH₂Cl₂ (2 mL). The reaction mixture wasstirred for 5 hours at room temperature and concentrated by removing thesolvent. 1 M hydrochloric acid (3 mL) and CH₂Cl₂ (2 mL) were added tothe residue. The resultant mixture was stirred for another 10 minutes atroom temperature. After removal of the supernatant, the solid was driedunder vacuum to afford the hydrochloride salt of compound 116 (93 mg) inan 80% yield.

CI-MS (M⁺+1): 520.4.

EXAMPLE 117 Preparation of Compound 117

Intermediate 117-I was obtained during the preparation of compound 62.

A solution of Intermediate 117-I (200 mg), cyclohexyl isocyanate (42mg), and Et₃N (62 mg) in CH₂Cl₂ (10 ml) was stirred at 25° C. forovernight and then concentrated. The resultant residue was purified bychromatography on silica gel (EtOAc/MeOH=20/1) to afford intermediate117-II (172 mg) in a 72% yield.

A solution of 20% TFA/CH₂Cl₂ (3 mL) was added to a solution of compound117-II (172 mg) in CH₂Cl₂ (2 mL). The reaction mixture was stirred for 5hours at room temperature and concentrated by removing the solvent. 1 Mhydrochloric acid (3 mL) and CH₂Cl₂ (2 mL) were added to the residue.The mixture was stirred for another 10 minutes at room temperature.After removal of the supernatant, the solid was dried under vacuum toafford the hydrochloride salt of compound 117 (145 mg) in a 91% yield.

CI-MS (M⁺+1): 577.4.

EXAMPLE 118 Preparation of Compound 118

Compound 118 was prepared in a manner similar to that used to preparecompound 117.

CI-MS (M⁺+1): 551.4.

EXAMPLE 119 Preparation of Compound 119

Compound 119 was prepared in a manner similar to that used to preparecompound 116.

CI-MS (M⁺+1): 565.4.

EXAMPLE 120 Preparation of Compound 120

Compound 120 was prepared in a manner similar to that used to preparecompound 117.

CI-MS (M⁺+1): 551.4.

EXAMPLE 121 Preparation of Compound 121

Intermediate 121-I was obtained during the preparation of compound 62.

A solution of intermediate 121-I (200 mg), trifluoroacetic anhydride(257 mg), and Et₃N (155 mg) in CH₂Cl₂ (10 ml) was stirred at 25° C. forovernight and then concentrated. The resultant residue was purified bychromatography on silica gel (EtOAc/MeOH=15/1) to afford intermediate121-II (163 mg) in a 71% yield.

A solution of 20% TFA/CH₂Cl₂ (3 mL) was added to a solution of compound121-II (163 mg) in CH₂Cl₂ (2 mL). The reaction mixture was stirred for 5hours at room temperature and concentrated by removing the solvent. 1 Mhydrochloric acid (3 mL) and CH₂Cl₂ (2 mL) were added to the residue.The mixture was stirred for another 10 minutes at room temperature.After removal of the supernatant, the solid was dried under vacuum toafford the hydrochloride salt of compound 121 (127 mg) in an 84% yield.

CI-MS (M⁺+1): 548.3.

EXAMPLE 122 Preparation of Compound 122

Compound 122 was prepared in a manner similar to that used to preparecompound 62.

CI-MS (M⁺+1): 600.4.

EXAMPLE 123 Preparation of Compound 123

Compound 123 was prepared in a manner similar to that used to preparecompound 62.

CI-MS (M⁺+1): 520.4.

EXAMPLE 124 Preparation of Compound 124

Compound 124 was prepared in a manner similar to that used to preparecompound 117.

CI-MS (M⁺+1): 567.4.

EXAMPLE 125 Preparation of Compound 125

Compound 125 was prepared in a manner similar to that used to preparecompound 117.

CI-MS (M⁺+1): 583.4.

EXAMPLE 126 Preparation of Compound 126

Compound 126 was prepared in a manner similar to that used to preparecompound 116.

CI-MS (M⁺+1): 522.4.

EXAMPLE 127 Preparation of Compound 127

Compound 127 was prepared in a manner similar to that used to preparecompound 116.

CI-MS (M⁺+1): 562.3.

EXAMPLE 128 Preparation of Compound 128

Compound 128 was prepared in a manner similar to that used to preparecompound 43.

CI-MS (M⁺+1): 514.3.

EXAMPLE 129 Preparation of Compound 129

Compound 129 was prepared in a manner similar to that used to preparecompound 43.

CI-MS (M⁺+1): 530.3.

EXAMPLE 130 Preparation of Compound 130

Compound 130 was prepared in a manner similar to that used to preparecompound 43.

CI-MS (M⁺+1): 528.3.

EXAMPLE 131 Preparation of Compound 131

Compound 131 was prepared in a manner similar to that used to preparecompound 43.

CI-MS (M⁺+1): 609.5.

EXAMPLE 132 Preparation of Compound 132

Compound 132 was prepared in a manner similar to that used to preparecompound 43.

CI-MS (M⁺+1): 613.4.

EXAMPLE 133 Preparation of Compound 133

Compound 133 was prepared in a manner similar to that used to preparecompound 43.

CI-MS (M⁺+1): 624.5.

EXAMPLE 134 Preparation of Compound 134

Compound 134 was prepared in a manner similar to that used to preparecompound 43.

CI-MS (M⁺+1): 622.4.

EXAMPLE 135 Preparation of Compound 135

Compound 135 was prepared in a manner similar to that used to preparecompound 109.

CI-MS (M⁺+1): 573.4.

EXAMPLE 136 Preparation of Compound 136

Compound 136 was prepared in a manner similar to that used to preparecompound 109.

CI-MS (M⁺+1): 599.3.

EXAMPLE 137 Preparation of Compound 137

Compound 137 was prepared in a manner similar to that used to preparecompound 21.

CI-MS (M⁺+1): 496.4.

EXAMPLE 138 Preparation of Compound 138

Compound 138 was prepared in a manner similar to that used to preparecompound 21.

CI-MS (M⁺+1): 511.4.

EXAMPLE 139 Preparation of Compound 139

Compound 139 was prepared in a manner similar to that used to preparecompound 21.

CI-MS (M⁺+1): 541.4.

EXAMPLE 140 Preparation of Compound 140

Compound 140 was prepared in a manner similar to that used to preparecompound 21.

CI-MS (M⁺+1): 510.4.

EXAMPLE 141 Preparation of Compound 141

Compound 141 was prepared in a manner similar to that used to preparecompound 44.

CI-MS (M⁺+1): 540.4.

EXAMPLE 142 Preparation of Compound 142

Intermediate 142-I was obtained during the preparation of compound 65.

A solution of intermediate 142-I (200 mg), methoxyacetyl chloride (60mg), and Et₃N (60 mg) in CH₂Cl₂ (10 ml) was stirred at 0° C. for 1 hourand then was allowed to warm-up to room temperature within 3 hours. Thesolution was then concentrated to give a residue, which was purified bychromatography on silica gel (EtOAc/Hexane=1/1) to afford intermediate142-II (107 g) in a 48% yield.

A solution of 20% TFA/CH₂Cl₂ (3 mL) was added to a solution of compound142-II (107 mg) in CH₂Cl₂ (2 mL). The reaction mixture was stirred for 5hours at room temperature and concentrated by removing the solvent. 1 Mhydrochloric acid (3 mL) and CH₂Cl₂ (2 mL) were added to the residue.The mixture was stirred for another 10 minutes at room temperature.After removal of the supernatant, the solid was dried under vacuum toafford the hydrochloride salt of compound 142 (92 mg) in a 93% yield.

CI-MS (M⁺+1): 597.4.

EXAMPLE 143 Preparation of Compound 143

Compound 143 was prepared in a manner similar to that used to preparecompound 142.

CI-MS (M⁺+1): 641.4.

EXAMPLE 144 Preparation of Compound 144

Acetyloxyacetyl chloride (304 mg) was added dropwise to a stirredsolution of compound 142-I (200 mg) and Et₃N (0.3 mL) in CH₂Cl₂ (10 mL)at 0° C. for 2 hours to afford a residue. The resultant residue waspurified by chromatography on silica gel (EtOAc/Hexane=1/1) to affordintermediate 144-II (200 mg) in a 90% yield.

An aqueous solution of 20% LiOH (4 mL) was added to 144-II (200 mg) inTHF (5 mL). After stirring for 12 hour, the mixture was acidified with2M HCl to obtain a crude product. The crude product was purified bychromatography on silica gel (EtOAc/MeOH=20/1) afforded intermediate144-III (98 mg) in a 51% yield.

Compound 144-III (98 mg) was treated with 20% TFA/CH₂Cl₂ (2 mL) at roomtemperature for 12 hours and then concentrated. The resultant residuewas purified by chromatography on silica gel (21% NH₃ (aq)/MeOH=1/19) toafford compound 144 (65 mg) in a 90% yield. Compound 144 was thentreated with 1 M HCl (2 mL) in CH₂Cl₂ (2 mL) for 0.5 hour. The solventswere evaporated and the residue was treated with ether and filtered toafford the hydrochloride salt of compound 144.

CI-MS (M⁺+1): 583.4.

EXAMPLE 145 Preparation of Compound 145

Compound 145 was prepared in the same manner as that used to prepareintermediate 144-II.

CI-MS (M⁺+1): 625.5.

EXAMPLE 146 Preparation of Compound 146

Compound 146 was prepared in a manner similar to that used to preparecompound 144.

CI-MS (M⁺+1): 625.5.

EXAMPLE 147 Preparation of Compound 147

Compound 147 was prepared in a manner similar to that used to preparecompound 44.

CI-MS (M⁺+1): 441.3.

EXAMPLE 148 Preparation of Compound 148

Compound 148 was prepared in a manner similar to that used to preparecompound 44.

CI-MS (M⁺+1): 654.0.

EXAMPLE 149 Preparation of Compound 149

Compound 149 was prepared in a manner similar to that used to preparecompound 116.

CI-MS (M⁺+1): 509.8.

EXAMPLE 150 Preparation of Compound 150

Compound 150 was prepared in a manner similar to that used to preparecompound 116.

CI-MS (M⁺+1): 567.7.

EXAMPLE 151 Preparation of Compound 151

Compound 151 was prepared in a manner similar to that used to preparecompound 116.

CI-MS (M⁺+1): 537.7.

EXAMPLE 152 Preparation of Compound 152

Compound 152 was prepared in a manner similar to that used to preparecompound 116.

CI-MS (M⁺+1): 620.3.

EXAMPLE 153 Preparation of Compound 153

Compound 153 was prepared in a manner similar to that used to preparecompound 116.

CI-MS (M⁺+1): 551.8.

EXAMPLE 154 Preparation of Compound 154

Compound 154 was prepared from compound 62-III in a manner similar tothat used to prepare compound 144.

CI-MS (M⁺+1): 538.4.

EXAMPLE 155 Preparation of Compound 155

Compound 155 was prepared from compound 62-III in a manner similar tothat used to prepare compound 144.

CI-MS (M⁺+1): 552.4.

EXAMPLE 156 Preparation of Compound 156

Compound 156 was prepared from compound 62-III in a manner similar tothat used to prepare compound 144.

CI-MS (M⁺+1): 510.3.

EXAMPLE 157 Preparation of Compound 157

Compound 157 was prepared from compound 109-I in a manner similar tothat used to prepare compound 109.

CI-MS (M⁺+1): 569.4.

EXAMPLE 158 Preparation of Compound 158

Compound 158 was prepared from compound 109-I in a manner similar tothat used to prepare compound 109.

CI-MS (M⁺+1): 539.4.

EXAMPLE 159 Preparation of Compound 159

Compound 159 was prepared from compound 109-I in a manner similar tothat used to prepare compound 109.

CI-MS (M⁺+1): 525.4.

EXAMPLE 160 Preparation of Compound 160

Compound 160 was prepared from compound 109-I in a manner similar tothat used to prepare compound 109.

CI-MS (M⁺+1): 567.4.

EXAMPLE 161 Preparation of Compound 161

Compound 161 was prepared from compound 21-II in a manner similar tothat used to prepare compound 154.

CI-MS (M⁺+1): 539.3.

EXAMPLE 162 Preparation of Compound 162

Compound 162 was prepared from compound 21-II in a manner similar tothat used to prepare compound 154.

CI-MS (M⁺+1): 511.3.

EXAMPLE 163 Preparation of Compound 163

Compound 163 was prepared from compound 21-II in a manner similar tothat used to prepare compound 154.

CI-MS (M⁺+1):511.1.

EXAMPLE 164 Preparation of Compound 164

Compound 164 was prepared from compound 21-II in a manner similar tothat used to prepare compound 154.

CI-MS (M⁺+1): 511.7.

EXAMPLE 165 Preparation of Compound 165

Compound 165 was prepared from compound 21-II in a manner similar tothat used to prepare compound 154.

CI-MS (M⁺+1): 539.4.

EXAMPLE 166 Preparation of Compound 166

Compound 166 was prepared in a manner similar to that used to preparecompound 43.

CI-MS (M⁺+1): 597.4.

EXAMPLE 167 Preparation of Compound 167

Compound 167 was prepared in a manner similar to that used to preparecompound 43.

CI-MS (M⁺+1): 613.4.

EXAMPLE 168 Preparation of Compound 168

Compound 168 was prepared in a manner similar to that used to preparecompound 109.

CI-MS (M⁺+1): 553.4.

EXAMPLE 169 Preparation of Compound 169

Compound 169 was prepared from compound 34-I in a manner similar to thatused to prepare compound 34.

CI-MS (M⁺+1): 520.4.

EXAMPLE 170 Preparation of Compound 170

Compound 170 was prepared from compound 34-I in a manner similar to thatused to prepare compound 34.

CI-MS (M⁺+1): 492.3.

EXAMPLE 171 Preparation of Compound 171

Compound 171 was prepared from compound 70-I and the correspondingthioisocyanate in a manner similar to that used to prepare compound 70.

CI-MS (M⁺+1): 570.3.

EXAMPLE 172 Preparation of Compound 172

Compound 172 was prepared from compound 70-I and the correspondingthioisocyanate in a manner similar to that used to prepare compound 70.

CI-MS (M⁺+1): 539.4.

EXAMPLE 173 Preparation of Compound 173

Compound 173 was prepared from compound 70-I and the correspondingthioisocyanate in a manner similar to that used to prepare compound 70.

CI-MS (M⁺+1): 574.3.

EXAMPLE 174 Preparation of Compound 174

Compound 174 was prepared from compound 70-I and the correspondingthioisocyanate in a manner similar to that used to prepare compound 70.

CI-MS (M⁺+1): 568.3.

EXAMPLE 175 Preparation of Compound 175

Compound 175 was prepared from compound 70-I and the correspondingthioisocyanate in a manner similar to that used to prepare compound 70.

CI-MS (M⁺+1): 553.4.

EXAMPLE 176 Preparation of Compound 176

Compound 176 was prepared from compound 70-I and the correspondingthioisocyanate in a manner similar to that used to prepare compound 70.

CI-MS (M⁺+1): 568.3.

EXAMPLE 177 Preparation of Compound 177

Compound 177 was prepared from compound 70-I and the correspondingthioisocyanate in a manner similar to that used to prepare compound 70.

CI-MS (M⁺+1): 552.2.

EXAMPLE 178 Preparation of Compound 178

Compound 178 was prepared from compound 70-I and the correspondingthioisocyanate in a manner similar to that used to prepare compound 70.

CI-MS (M⁺+1): 554.2.

EXAMPLE 179 Preparation of Compound 179

Compound 179 was prepared from compound 70-I and the correspondingthioisocyanate in a manner similar to that used to prepare compound 70.

CI-MS (M⁺+1): 584.3.

EXAMPLE 180 Preparation of Compound 180

Compound 180 was prepared from compound 70-I and the correspondingthioisocyanate in a manner similar to that used to prepare compound 70.

CI-MS (M⁺+1): 598.5.

EXAMPLE 181 Preparation of Compound 181

Compound 181 was prepared from compound 70-I and the correspondingthioisocyanate in a manner similar to that used to prepare compound 70.

CI-MS (M⁺+1): 553.4.

EXAMPLE 182 Preparation of Compound 182

Compound 182 was prepared from compound 109-I in a manner similar tothat used to prepare compound 109.

CI-MS (M⁺+1): 566.3.

EXAMPLE 183 Preparation of Compound 183

Compound 183 was prepared from compound 109-I in a manner similar tothat used to prepare compound 109.

CI-MS (M⁺+1): 524.3.

EXAMPLE 184 Preparation of Compound 184

Compound 184 was prepared from compound 109-I in a manner similar tothat used to prepare compound 109.

CI-MS (M⁺+1): 565.3.

EXAMPLE 185 Preparation of Compound 185

Compound 185 was prepared from compound 109-I in a manner similar tothat used to prepare compound 109.

CI-MS (M⁺+1): 550.3.

EXAMPLE 186 Preparation of Compound 186

Compound 186 was prepared from compound 109-I in a manner similar tothat used to prepare compound 109.

CI-MS (M⁺+1): 547.4.

EXAMPLE 187 Preparation of Compound 187

Compound 187 was prepared from compound 35-I in a manner similar to thatused to prepare compound 35.

CI-MS (M⁺+1): 533.4.

EXAMPLE 188 Preparation of Compound 188

Compound 188 was prepared from compound 33-I in a manner similar to thatused to prepare compound 33.

CI-MS (M⁺+1): 537.4.

EXAMPLE 189 Preparation of Compound 189

Compound 189 was prepared from compound 33-I in a manner similar to thatused to prepare compound 33.

CI-MS (M⁺+1): 583.4.

EXAMPLE 190 Preparation of Compound 190

Compound 190 was prepared from compound 33-I in a manner similar to thatused to prepare compound 33.

CI-MS (M⁺+1): 537.4.

EXAMPLE 191 Preparation of Compound 191

Compound 191 was prepared in a manner similar to that used to preparecompound 44.

CI-MS (M⁺+1): 463.2.

EXAMPLE 192 Preparation of Compound 192

tert-Butoxycarbonylaminoethylamine (5.4 g) was added to a solution of2,4-dichloro-6-methylpyrimidine (5 g) in THF (350 mL). The reactionmixture was stirred at 0° C. for 1 hour and then was allowed to warm-upto room temperature within 3 hours. After the solution was thenconcentrated and treated with 1 M HCl (40 mL) in MeOH (50 mL), it wasstirred at room temperature for another 8 hours. The solution was thenagain concentrated. The resultant residue was then neutralization withNH₄OH and extracted with CH₂Cl₂. The solution was concentrated and theresidue was purified by chromatography on silica gel (MeOH as eluant) toafford intermediate 192-II (4.6 g) in a 90% yield.

Butyryl chloride (430 mg) was added to a solution of 192-II (680 mg) inCH₂Cl₂ (35 mL). After 1 hour of stirring at room temperature, thesolution was concentrated and 1-pentanol (2 mL) was added.Diisopropylethylamine (0.2 mL), intermediate 8-III prepared in example 8(150 mg), and NaI (110 mg) were also added to this solution, which wasthen stirred for 24 hours at 120° C. The resultant mixture was dissolvedin CH₂Cl₂, washed with water, dried with anhydrous MgSO₄ and evaporatedto afford 192-IV in a 50% yield after purification by chromatography onsilica gel (EtOAc/MeOH=5/1).

192-IV (950 mg) was treated with 1 M HCl (20 mL) and stirred forovernight. After the supernatant was removed, compound 192 was collectedby filtration.

CI-MS (M⁺+1): 498.4.

EXAMPLE 193 Preparation of Compound 193

Compound 193 was prepared in a manner similar to that used to preparecompound 192.

CI-MS (M⁺+1): 493.4.

EXAMPLE 194 Preparation of Compound 194

Compound 194 was prepared in a manner similar to that used to preparecompound 192.

CI-MS (M⁺+1): 542.4.

EXAMPLE 195 Preparation of Compound 195

Compound 195 was prepared in a manner similar to that used to preparecompound 192.

CI-MS (M⁺+1): 510.4.

EXAMPLE 196 Preparation of Compound 196

Compound 196 was prepared in a manner similar to that used to preparecompound 192.

CI-MS (M⁺+1): 526.4.

EXAMPLE 197 Preparation of Compound 197

Compound 197 was prepared in a manner similar to that used to preparecompound 192.

CI-MS (M⁺+1): 524.4.

EXAMPLE 198 Preparation of Compound 198

2,4-Dichloro-6-aminopyrimidine (2.0 g) was dissolved in 1-pentanol (10mL). Cyclohexylaminopropylamine (1.92 g) was then added. The reactionmixture was stirred at 120° C. for 24 hours. The solution wasconcentrated and the residue was purified by column chromatography onsilica gel (EtOAc/Hexane=1/3) to afford 198-I (1.8 mg) in a 52% yield.

A solution of intermediate 198-I (1.8 g) reacted with (Boc)₂O in CH₂Cl₂(120 mL) for 8 hours at 25° C. The solution was concentrated and theresidue was purified by column chromatography on silica gel(EtOAc/Hexane=1/9) to give pure 198-II (1.06 g) in a 70% yield.

N¹-Morpholine-N¹-piperazine ethane (0.3 g) was added to 198-II (130 mg).The mixture was stirred at 120° C. for 8 hours. The solution wasconcentrated and the residue was treated with water and extracted withCH₂Cl₂. The organic layer was collected, concentrated to give a crudeproduct, which was purified by column chromatography on silica gel(EtOAc/MeOH=10/1) to afford 198-III (100 mg) in a 72% yield.

Compound 198-III (100 mg) was treated with 20% TFA/CH₂Cl₂ (2 mL) for 8hours and then concentrated. The resultant residue was purified bychromatography on silica gel (21% NH₃ (aq)/MeOH=1/19) to afford Compound198 (69 mg) in a 85% yield. Compound 198 was then treated with 1 M HCl(2 mL) in CH₂Cl₂ (2 mL) for 0.5 hour. After the solvents wereevaporated, the residue was treated with ether and filtered to givehydrochloride salt of 198.

CI-MS (M⁺+1): 447.4.

EXAMPLE 199 Preparation of Compound 199

Compound 199 was prepared in a manner similar to that used to preparecompound 197.

CI-MS (M⁺+1): 408.3.

EXAMPLE 200 Preparation of Compound 200

Compound 200 was prepared in a manner similar to that used to preparecompound 197.

CI-MS (M⁺+1): 422.2.

EXAMPLE 201 Preparation of Compound 201

A solution of 3-cyanobenzylaldehyde (1.0 g) andN-cyclohexyl-1,3-propane-diamine (2.4 g) in CH₃OH (150 mL) was heated to60° C. for 18 hours. After cooling to room temperature, NaBH₄ (1.5 g)was slowly added to the above solution. The mixture was stirred foranother 30 minutes. The mixture was then concentrated, quenched withNH₄Cl (aq), and extracted with CH₂Cl₂. The organic layers were combined,dried with anhydrous MgSO₄, and concentrated to give a residue. Theresidue was purified by chromatography on silica gel (EtOAc/Et₃N=7/3) toafford Intermediate 201-I (1.6 g) in a 80% yield.

A solution of Intermediate 201-I (1.6 g) and Boc₂O (3.5 g) in CH₂Cl₂(160 ml) was stirred at 25° C. for 15 hours and then concentrated. Theresultant residue was purified by chromatography on silica gel(EtOAc/Hexane=1/1) to afford Intermediate 201-II as a yellow oil (2.36g) in a 85% yield.

A solution of Intermediate 201-II and LiAlH₄ (2.3 g) in THF (230 mL) wasstirred at 0° C. for 4 hours. After Na₂SO₄.10H₂O was added, the solutionwas stirred at room temperature for 0.5 hour. The solution was thenfiltered through a celite pad. The filtrate was dried over anhydrousMgSO₄ and concentrated to give a residue. The residue was purified bycolumn chromatography on silica gel (using MeOH as an eluant) to affordIntermediate 201-III (1.1 g) in a 50% yield.

Diisopropylethylamine (1.1 mL) was added to a solution of2,4-dichloro-6-aminopiperidine (0.41 g) and Intermediate 201-III (1.1 g)in 1-pentanol (10 mL). The reaction mixture was stirred overnight at120° C. The solvent was removed under vacuum and the residue waspurified by column chromatography on silica gel (EtOAc/Hexane=3/7) toafford 201-IV (1.0 g) in a 65% yield.

To a solution of Intermediate 201-IV (1.0 g) in 1-pentanol (1 mL) wasadded N¹-hydroxyethoxyethyl piperazine (0.25 g). After the solution wasstirred at 120° C. for 8 hours, it was concentrated. The residue thusobtained was purified by column chromatography on silica gel(EtOAc/MeOH=4/1) to afford Intermediate 201-V (730 mg) in a 60% yield.

A solution of 20% TFA/CH₂Cl₂ (5 mL) was added to a solution ofIntermediate 201-V (0.73 g) in CH₂Cl₂ (2 mL). The reaction mixture wasstirred for 5 hours at room temperature and concentrated by removing thesolvent. The resultant residue was purified by column chromatography onsilica gel (21% NH₃ (aq)/MeOH=1/19) to afford Compound 201 (434 mg) in a85% yield. Compound 201 was then treated with 1 M HCl (4 mL) in CH₂Cl₂(2 mL) for 0.5 hour. After the solvents were removed, the residue wastreated with ether and filtered to give hydrochloride salt of compound201.

CI-MS (M⁺+1): 541.3.

EXAMPLE 202 Preparation of Compound 202

Compound 202 was prepared in a manner similar to that used to preparecompound 200.

CI-MS (M⁺+1): 566.4.

EXAMPLE 203 Preparation of Compound 203

Compound 203 was prepared in a manner similar to that used to preparecompound 200.

CI-MS (M⁺+1): 525.4.

EXAMPLE 204 Preparation of Compound 204

A solution of 4-cyanobenzaldehyde (3.0 g) andtert-butoxyaminopropylamine (3.9 g) in MeOH (60 mL) was heated at 60° C.for 6 hours. After the solution was cooled to room temperature, NaBH₄(2.5 g) was slowly added. After the mixture was stirred for 30 minutes,it was concentrated, quenched with NH₄Cl (aq.), and extracted withCH₂Cl₂. The organic layer was separated and concentrated. The residuethus obtained was purified by column chromatography on silica gel(EtOAc/Hexane=1/3) to afford Intermediate 204-I (5.8 g) in a 88% yield.

A solution of intermediate 204-I (5.8 g) and 1 M HCl (40 mL) in MeOH (50mL) was stirred at room temperature for 8 hours. The solution was thenconcentrated and the resultant residue was neutralization with NH₄OH,and extracted with CH₂Cl₂. The organic layer was separated andconcentrated. The residue thus obtained was purified by columnchromatography on silica gel (using MeOH as an eluant) to affordIntermediate 204-II (3.4 g) in a 90% yield.

To a solution of compound 204-II (3.5 g) in MeOH (50 mL) was addedtetrahydro-4H-pyran-4-one (2 g). The solution was then heated at 60° C.for 6 hours. After the solution was cooled to room temperature, NaBH₄(1.85 g) was slowly added. After the mixture was stirred 30 minutes, itwas concentrated, quenched with NH₄Cl (aq.), and extracted with CH₂Cl₂.The organic layer was separated and concentrated. The residue thusobtained was purified by column chromatography on silica gel(EtOAc/MeOH=1/1) to afford Intermediate 204-III (3.5 g) in a 70% yield.

A solution of Intermediate 204-III (12.99 g) and Boc₂O (20.76 g) in1,4-dioxane (200 ml) and H₂O (100 mL) was stirred at room temperaturefor 15 hours and then concentrated. The resultant residue was purifiedby column chromatography on silica gel (EtOAc/Hexane=1/3) to affordIntermediate 204-IV (21.5 g) in a 95% yield.

LiAlH4 (8.6 g) was added to a solution of Intermediate 204-IV (21.5 g)in THF (500 mL) and ether (500 mL). After the solution was stirred at 0°C. for 2 hours, it was treated with saturated aq. NH₄Cl solution,extracted with CH₂Cl₂, and concentrated. The residue thus obtained waspurified by column chromatography on silica gel (using MeOH as aneluant) to afford Intermediate 204-V (13.0 g) in a 60% yield.

A solution of Intermediate 204-V (7.6 g) in 1-pentanol (50 mL) wasreacted with 2,4-dichloro-6-aminopyrimidine (3.1 g) at 120° C. for 12 h.The solvent was then removed and the residue was purified by columnchromatography on silica gel (EtOAc/MeOH=5/1) to afford Intermediate204-VI (7.2 g) in a 75% yield.

Intermediate 204-VI (400 mg) was added to N¹-Morpholine-N¹-piperazineethane (470 mg) in 1-pentanol (1 mL). The reaction mixture was heated at120° C. for 12 hours. The solvent was then removed under vacuum and theresidue was purified by column chromatography on silica gel(EtOAc/MeOH=1/1) to afford Intermediate 204-VII (386 mg) in a 76% yield.

A solution of 20% TFA/CH₂Cl₂ (3 mL) was added to a solution ofIntermediate 204-VI (386 mg) in CH₂Cl₂ (2 mL). The reaction mixture wasstirred for 8 hours at room temperature and concentrated by removing thesolvent. The resultant residue was purified by column chromatography onsilica gel (21% NH₃ (aq)/MeOH=1/19) to afford Compound 204 (256 mg) in a85% yield. Compound 204 was then treated with 1 M HCl (3 mL) in CH₂Cl₂(1 mL) for 0.5 hour. After the solvents were removed, the residue wastreated with ether and filtered to give hydrochloride salt of 204.

CI-MS (M⁺+1): 568.4.

EXAMPLE 205 Preparation of Compound 205

Compound 205 was prepared in a manner similar to that used to preparecompound 204.

CI-MS (M⁺+1): 527.3.

EXAMPLE 206 Preparation of Compound 206

Compound 206 was prepared in a manner similar to that used to preparecompound 204.

CI-MS (M⁺+1): 543.3.

EXAMPLE 207 Preparation of Compound 207

Compound 207 was prepared in a manner similar to that used to preparecompound 204.

CI-MS (M⁺+1): 554.4.

EXAMPLE 208 Preparation of Compound 208

Compound 208 was prepared in a manner similar to that used to preparecompound 204.

CI-MS (M⁺+1): 529.3.

EXAMPLE 209 Preparation of Compound 209

Compound 209 was prepared in a manner similar to that used to preparecompound 204.

CI-MS (M⁺+1): 513.3.

EXAMPLE 210 Preparation of Compound 210

Compound 210 was prepared in a manner similar to that used to preparecompound 204.

CI-MS (M⁺+1): 538.4.

EXAMPLE 211 Preparation of Compound 211

Compound 211 was prepared in a manner similar to that used to preparecompound 204.

CI-MS (M⁺+1): 513.4.

EXAMPLE 212 Preparation of Compound 212

Compound 212 was prepared in a manner similar to that used to preparecompound 204.

CI-MS (M⁺+1): 497.3.

EXAMPLE 213 Preparation of Compound 213

Compound 213 was prepared in a manner similar to that used to preparecompound 204.

CI-MS (M⁺+1): 512.4.

EXAMPLE 214 Preparation of Compound 214

Compound 214 was prepared in a manner similar to that used to preparecompound 204.

CI-MS (M⁺+1): 471.4.

EXAMPLE 215 Preparation of Compound 215

Compound 215 was prepared in a manner similar to that used to preparecompound 204.

CI-MS (M⁺+1): 501.4.

EXAMPLE 216 Preparation of Compound 216

Compound 216 was prepared in a manner similar to that used to preparecompound 204.

CI-MS (M⁺+1): 485.3.

EXAMPLE 217 Preparation of Compound 217

Compound 217 was prepared in a manner similar to that used to preparecompound 204.

CI-MS (M⁺+1): 578.4.

EXAMPLE 218 Preparation of Compound 218

Compound 218 was prepared in a manner similar to that used to preparecompound 204.

CI-MS (M⁺+1): 553.4.

EXAMPLE 219 Preparation of Compound 219

Compound 219 was prepared in a manner similar to that used to preparecompound 204.

CI-MS (M⁺+1): 537.4.

EXAMPLE 220 Preparation of Compound 220

Compound 220 was prepared in a manner similar to that used to preparecompound 204.

CI-MS (M⁺+1): 553.3.

EXAMPLE 221 Preparation of Compound 221

Compound 221 was prepared in a manner similar to that used to preparecompound 204.

CI-MS (M⁺+1): 537.4.

EXAMPLE 222 Preparation of Compound 222

NaBH(OAc)₃ (52.68 g) was added to a solution of ethyl (2-aminomethyl)acetate (25.5 g) and cyclohexanone (24.45 g) in CH₂Cl₂ (200 mL) and MeOH(300 ml). The mixture was stirred at room temperature for 6 hours. Aftera saturated aq. NaHCO₃ solution was added, the mixture was extractedwith EtOAc, dried over anhydrous MgSO₄, and filtered. The solvent wasthen removed and the residue was purified by column chromatography onsilica gel (EtOAc/Hexane=1/1) to afford Intermediate 222-I (32.9 g) in a76% yield.

A solution of Intermediate 222-I (32.9 g) and Boc₂O (36.0 g) in CH₂Cl₂(300 mL) was stirred at 25° C. overnight. The solution was thenconcentrated and the resultant residue was purified by columnchromatography on silica gel (EtOAc/Hexane=1/9) to give Intermediate222-II (39.5 g) in a 80% yield.

1M DIBAL/ether (85 mL) was added to a stirred solution of Intermediate222-II (15 g) in dry toluene (500 mL) at −70˜−78° C. under N₂ (g). Thereaction mixture was stirred for 2 hours at this temperature. After 5%HCl (aq) (85 mL) was then added to the solution at −60˜−70° C., themixture was stirred for another 0.5 hour after the reaction temperaturewas increased to 25° C. The aqueous layer was extracted with CH₂Cl₂twice. The organic layers were combined, dried with anhydrous MgSO₄, andconcentrated by removing the solvent under vacuum. The resultant residuewas purified by column chromatography on silica gel (EtOAc/Hexane=1/5)to afford Intermediate 222-III (7.7 g) in a 60% yield.

Di-2-aminoethylether (1.0 g) was slowly added to a stirred solution of2,4-dichloro-6-methylpyrimidine (2.0 g) in THF (15 mL) at roomtemperature. The mixture was stirred at 0° C. for 2 hours and thereaction was allowed to warm-up to room temperature overnight. Thesolution was then concentrated and the resultant residue was purified bycolumn chromatography on silica gel (EtOAc/MeOH=1/1) to giveIntermediate 222-IV (1.5 g) in a 53% yield.

A solution of Intermediate 222-IV (0.7 g) and Intermediate 222-III (0.77g) in MeOH (60 mL) was stirred at 60° C. for 8 hours. NaBH₄ (0.17 g) wasthen added the solution at 0° C. After the solution was stirred for 0.5hour, a saturated aq. NH₄Cl solution was added and the mixture wasextracted with EtOAc, dried over anhydrous MgSO₄, and filtered. Thesolution was then concentrated and the resultant residue was purified bycolumn chromatography on silica gel (EtOAc/MeOH=2/1) to affordIntermediate 222-V (356 mg) in a 25% yield.

A solution of Intermediate 222-V (356 mg) and Boc₂O (180 mg) in CH₂Cl₂(10 mL) was stirred at 25° C. overnight. The solution was thenconcentrated and the resultant residue was purified by columnchromatography on silica gel (EtOAc/Hexane=1/5) to give Intermediate222-VI (410 mg) in a 95% yield.

N¹-Morpholine-N¹-piperazine ethane (221 mg) was added to a solution ofIntermediate 222-VI (210 mg) in 1-pentanol (1 mL). The mixture wasstirred at 120° C. for 12 hours. It was then concentrated and theresultant residue was purified by column chromatography on silica gel(EtOAc/MeOH=10/1) to afford Intermediate 222-VII (100 mg) in a 37%yield.

A solution of 20% TFA/CH₂Cl₂ (2 mL) was added to a solution ofIntermediate 222-VII (100 mg) in CH₂Cl₂ (1 mL). The reaction mixture wasstirred for 8 hours at room temperature and concentrated by removing thesolvent. The resultant residue was purified by column chromatography onsilica gel (21% NH₃ (aq)/MeOH=1/19) to afford Compound 222 (65 mg) in90% yield. Compound 222 was then treated with 1 M HCl (2 mL) in CH₂Cl₂(1 mL) for 0.5 hour. After the solvents were removed, the residue wastreated with ether and filtered to afford hydrochloride salt of 222.

CI-MS (M⁺+1): 533.4.

EXAMPLE 223 Preparation of Compound 223

Compound 223 was prepared in a manner similar to that used to preparecompound 222.

CI-MS (M⁺+1): 528.4.

EXAMPLE 224 Preparation of Compound 224

N-tert-Butoxycarbonylpiperidinyl-4-methylamine (5.0 g) was slowly addedto a stirred solution of 2,4-dichloro-6-aminopyrimidine (5.7 g) in1-pentanol (20 mL). The solution was stirred at 120° C. for 12 hours.The solution was then concentrated and the resultant residue waspurified by column chromatography on silica gel (EtOAc/Hexane=1/9) togive Intermediate 224-I (3.6 g) in a 45% yield.

Intermediate 224-I (2.4 g) was then dissolved in CH₂Cl₂ (80 mL) and 20%TFA/CH₂Cl₂ (20 mL) was added. The solution was stirred at roomtemperature overnight. The solution was then concentrated and theresultant residue was purified by column chromatography on silica gel(21% NH₃ (aq)/MeOH=1/19) to afford Intermediate 224-II (1.5 g) in a 90%yield.

Intermediate 222-III (3.3 g) prepared in Example 222 was added to asolution of Intermediate 224-II (1.9 g) in MeOH (40 mL). The mixture wasstirred at 60° C. for 12 hours. NaBH₄ (0.3 g) was then added at 0° C.After the mixture was stirred for 1 hour, an aqueous solution of NH₄Cl(10%, 10 mL) was added. The mixture was extracted with EtOAc, dried overanhydrous MgSO₄, and filtered. The solution thus obtained was thenconcentrated. The resultant residue was purified by columnchromatography on silica gel (EtOAc/Hexane=1/1) to afford Intermediate224-III (1.5 g) in a 40% yield.

N¹-Morpholine-N¹-piperazine ethane (370 mg) was added to Intermediate224-III (300 mg) in 1-pentanol (1 mL). The mixture was stirred at 120°C. for 12 hours. After the solution was concentrated, the residue wastreated with water and extracted with CH₂Cl₂. The organic layer wasseparated and concentrated. The resultant residue was purified by columnchromatography on silica gel (EtOAc/Hexane=1/9) to afford Intermediate224-IV (281 mg) in a 70% yield.

A solution of 20% TFA/CH₂Cl₂ (3 mL) was added to a solution ofIntermediate 224-IV (281 mg) in CH₂Cl₂ (2 mL). The reaction mixture wasstirred for 8 hours at room temperature and concentrated by removing thesolvent. The resultant residue was purified by column chromatography onsilica gel (21% NH₃ (aq)/MeOH=1/19) to afford Compound 224 (200 mg) in a85% yield. Compound 224 was then treated with 1 M HCl (4 mL) in CH₂Cl₂(2 mL) for 0.5 hours. After the solvents were removed, the residue wastreated with ether and filtered to give hydrochloride salt of Compound224.

CI-MS (M⁺+1): 544.4.

EXAMPLE 225 Preparation of Compound 225

Compound 225 was prepared in a manner similar to that used to preparecompound 224.

CI-MS (M⁺+1): 503.4.

EXAMPLE 226 Preparation of Compound 226

Compound 226 was prepared in a manner similar to that used to preparecompound 224.

CI-MS (M⁺+1): 519.4.

EXAMPLE 227 Preparation of Compound 227

A solution of piperidinyl-4-methylamine (3.6 g) andN-tert-butoxycarbonylimidazole (5.3 g) in toluene (80 mL) was stirred at25° C. overnight. The solution was then concentrated and the resultantresidue was purified by column chromatography on silica gel(EtOAc/Hexane=1/2) to give Intermediate 227-I (4.7 g) in a 70% yield.

Intermediate 227-I (4.7 g) and Et₃N (2.7 mL) in 1-pentanol (20 mL) wasreacted with 2,4-dichloro-6-aminopyrimidine (5.4 g) at 120° C. for 12hours. After the solvent was removed, the residue was purified by columnchromatography on silica gel (EtOAc/Hexane=1/9) to afford Intermediate227-II (5.2 g) in a 70% yield.

A solution of Intermediate 227-II (1.0 g) treated with 1 M HCl (20 mL)in CH₂Cl₂ (10 mL) was stirred at room temperature for 8 hours. After thesolution was concentrated, the resultant residue was neutralization withNH₄OH, and extracted with CH₂Cl₂. The organic layer was separated andconcentrated. The residue thus obtained was purified by columnchromatography on silica gel (using MeOH as an eluant) to affordIntermediate 227-III (636 mg) in a 90% yield.

Intermediate 222-III (790 mg) prepared from Example 222 was added to asolution of Intermediate 227-III (450 mg) in MeOH (20 mL). The mixturewas stirred at 25° C. for 2 hours. NaBH(OAc)₃ (2.0 g) was then added at25° C. for 12 hours. After the solution was concentrated, a saturatedaq. NaHCO₃ solution was added to the resultant residue. The mixture wasthen extracted with CH₂Cl₂. The organic layer was separated andconcentrated. The residue thus obtained was purified by columnchromatography on silica gel (using MeOH as an eluant) to affordIntermediate 227-IV (539 mg) in a 60% yield.

N¹-Morpholine-N¹-piperazine ethane (240 mg) was added to a solution ofIntermediate 227-IV (160 mg) in 1-pentanol (1 mL). The mixture wasstirred at 120° C. for 8 hours. The solution was concentrated and theresidue was purified by column chromatography on silica gel(EtOAc/MeOH=5/1) to afford Intermediate 227-V (85 mg) in a 40% yield.

A solution of 20% TFA/CH₂Cl₂ (1 mL) was added to a solution ofIntermediate 227-V (85 mg) in CH₂Cl₂ (1 mL). The reaction mixture wasstirred for 8 hours at room temperature and concentrated by removing thesolvent. The resultant residue was purified by column chromatography onsilica gel (21% NH₃ (aq)/MeOH=1/19) to afford Compound 227 (65 mg) in a90% yield. Compound 227 was then treated with 1 M HCl (1 mL) in CH₂Cl₂(1 mL) for 0.5 hour. After the solvents were removed, the residue wastreated with ether and filtered to give hydrochloride salt of Compound227.

CI-MS (M⁺+1): 544.4.

EXAMPLE 228 Preparation of Compound 228

To a solution of 22-II (500 mg) in THF (10 mL) was added sulfur trioxidepyridine complex (457 mg). The mixture was stirred at 25° C. for 12hours. The solution was filtered and concentrated. The resultant residuewas purified by column chromatography on silica gel (EA/MeOH=10/1) togive Intermediate 228-II (82 mg) in a 10% yield.

A solution of 20% TFA/CH₂Cl₂ (1 mL) was added to a solution ofIntermediate 228-II (82 mg) in CH₂Cl₂ (1 mL). The reaction mixture wasstirred for 8 hours at room temperature and concentrated by removing thesolvent. The resultant residue was purified by chromatography on silicagel (21% NH₃ (aq)/MeOH=1/19) to afford Compound 228 (54 mg) in 90%yield. Compound 228 was then treated with 1 M HCl (1 mL) in CH₂Cl₂ (1mL) for 0.5 hour. After the solvents were removed, the residue wastreated with ether and filtered to give hydrochloride salt of Compound228.

CI-MS (M⁺+1): 577.2.

EXAMPLE 229 Preparation of Compound 229

Diethyl vinylphosphonate (377 mg) was added to a solution ofIntermediate 21-II (500 mg) prepared from Example 21 in MeOH (10 mL).The solution was stirred at 25° C. for 12 hours. The solution wasconcentrated and the residue was purified by column chromatography onsilica gel (EA/MeOH=5/1) to afford Intermediate 229-II (438 mg) in a 70%yield.

A solution of 20% TFA/CH₂Cl₂ (5 mL) was added to a solution ofIntermediate 229-II (438 mg) in CH₂Cl₂ (2 mL). The reaction mixture wasstirred for 8 hours at room temperature and concentrated by removing thesolvent. The resultant residue was purified by column chromatography onsilica gel (EA/MeOH=1/1) to afford Compound 229 (165 mg) in a 50% yield.

CI-MS (M⁺+1): 617.4.

EXAMPLE 230 Preparation of Compound 230

A solution of Compound 229 (600 mg) and trimethylsilyl bromide (1.19 g)in CH₂Cl₂ (30 mL) was stirred at 25° C. for 72 hours. The solution wasthen concentrated in vacuo to yield a yellow-orange foam, which wasre-dissolved in water (50 mL). The solution was washed with ether (3×35mL) and then concentrated in vacuo to yield a solid. The solid waspurified through a column of cation exchange resin (Dowex AG50X8) byeluting the column first with water (ca. 500 mL), and then with 0.2 Maqueous ammonia to afford ammonium salt of Compound 230 (58 mg) in a 10%yield.

CI-MS (M⁺+1): 561.1.

EXAMPLE 231 Preparation of Compound 231

Intermediate 222-III (4.5 g) was added to a solution oftrans-1,4-diaminocyclohexane (3 g) in MeOH (200 mL). The mixture wasstirred at 60° C. for 8 hours. After NaBH₄ (0.7 g) was added at 0° C.,the mixture was stirred for 0.5 hour and then concentrated by removingthe solvent. An aqueous solution of NH₄Cl (10%, 10 mL) was added to theresultant residue. The mixture was extracted with CH₂Cl₂, dried overanhydrous MgSO₄, filtered, and concentrated. The residue was purified bycolumn chromatography on silica gel (using MeOH as an eluant) to affordIntermediate 231-I (6.0 g) in a 65% yield.

Intermediate 231-I (6.0 g) and Et₃N (6.0 mL) in 1-pentanol (30 mL) wasreacted with 2,4-dichloro-6-aminopyrimidine (2.7 g) at 120° C. for 12hours. The solvent was then removed and the residue was purified bycolumn chromatography on silica gel (EtOAc/Hexane=1/1) to affordIntermediate 231-II (5.7 g) in a 70% yield.

N¹-Morpholine-N¹-piperazine ethane (620 mg) was added to Intermediate231-II (500 mg) in 1-pentanol (5 mL). The mixture was stirred at 120° C.for 8 hours and then concentrated. The residue thus obtained was treatedwith water and extracted with CH₂Cl₂ to afford Intermediate 231-III (468mg) in a 70% yield, which was purified by column chromatography onsilica gel using 21% NH₃ (aq) and MeOH as eluants.

A solution of 20% TFA/CH₂Cl₂ (5 mL) was added to a solution ofIntermediate 231-III (468 mg) in CH₂Cl₂ (2 mL). The reaction mixture wasstirred for 8 hours at room temperature and concentrated by removing thesolvent. The resultant residue was purified by column chromatography onsilica gel (21% NH₃ (aq)/MeOH=1/19) to afford Compound 231 (356 mg) in a90% yield. Compound 231 was then treated with 1 M HCl (4 mL) in CH₂Cl₂(2 mL) for 0.5 hour. After the solvents were removed, the residue wastreated with ether and filtered to give hydrochloride salt of Compound231.

CI-MS (M⁺+1): 544.4.

EXAMPLE 232 Preparation of Compound 232

Compound 232 was prepared in a manner similar to that used to preparecompound 231.

CI-MS (M⁺+1): 503.4.

EXAMPLE 233 Preparation of Compound 233

Compound 233 was prepared in a manner similar to that used to preparecompound 231.

CI-MS (M⁺+1): 519.4.

EXAMPLE 234 Preparation of Compound 234

Compound 8-III (1.0 g) prepared in Example 8 in MeOH (20 mL) washydrogenated in the presence of 10% Pd/C (200 mg) at 50 psi at roomtemperature for 18 hours. The mixture was then filtered and concentratedto afford Intermediate 234-II (500 mg) without further purification.

Crude Intermediate 234-II (0.5 g) in 1-pentanol (3 mL) was reacted with2,4-dichloro-6-aminopyrimidine (0.2 g) at 120° C. for 15 hours. Thesolution was then concentrated and the resultant residue was purified bycolumn chromatography on silica gel (EtOAc/Hexane=3/7) to giveIntermediate 234-III (0.3 g) in a 65% yield.

N¹-Morpholine-N¹-piperazine ethane (0.3 g) was added to Intermediate234-III (0.5 g) in 1-pentanol (1 mL). The mixture was stirred at 120° C.for 18 hours. The solution was concentrated to give the residue, whichwas then coated with SiO₂ and purified by column chromatography onsilica gel (EtOAc/MeOH=7/3) to afford Intermediate 234-IV (0.23 g) in a60% yield.

A solution of 20% TFA/CH₂Cl₂ (5 mL) was added to a solution ofIntermediate 234-IV (230 mg) in CH₂Cl₂ (2 mL). The reaction mixture wasstirred for 8 hours at room temperature and concentrated by removing thesolvent. The resultant residue was purified by column chromatography onsilica gel (21% NH₃ (aq)/MeOH=1/19) to afford Compound 234 (192 mg) in a85% yield. Compound 234 was then treated with 1 M HCl (4 mL) in CH₂Cl₂(2 mL) for 0.5 hour. After the solvents were removed, the residue wastreated with ether and filtered to give hydrochloride salt of Compound234.

CI-MS (M⁺+1): 572.5.

EXAMPLE 235 Preparation of Compound 235

Compound 235 was prepared in a manner similar to that used to preparecompound 234.

CI-MS (M⁺+1): 531.4.

EXAMPLE 236 Preparation of Compound 236

Compound 236 was prepared in a manner similar to that used to preparecompound 234.

CI-MS (M⁺+1): 547.4.

EXAMPLE 237 Preparation of Compound 237

Compound 237 was prepared in a manner similar to that used to preparecompound 234.

CI-MS (M⁺+1): 555.5.

EXAMPLE 238 Preparation of Compound 238

Compound 238 was prepared in a manner similar to that used to preparecompound 234.

CI-MS (M⁺+1): 549.4.

EXAMPLE 239 Preparation of Compound 239

Compound 239 was prepared in a manner similar to that used to preparecompound 234.

CI-MS (M⁺+1): 503.4.

EXAMPLE 240 Preparation of Compound 240

Intermediate 234-III (1.0 g) prepared from Example 3 was added to astirred solution of piperazine (0.36 g) in 1-pentanol (1.0 mL). Thesolution was stirred at 120° C. for 18 hours. It was concentrated togive a residue, which was coated with SiO₂ and purified by columnchromatography on silica gel (EtOAc/MeOH=9/1) to afford Intermediate240-I (0.82 g) in a 75% yield.

Methoxycarbonylacetyl chloride (0.2 g) was added to a solution ofIntermediate 240-I (0.82 g) in CH₂Cl₂ (50 mL) and Et₃N (0.3 g) at 0° C.The mixture was stirred at 0° C. for 1 hour and then concentrated. Theresidue thus obtained was treated with water and extracted with CH₂Cl₂.The organic layer was separated and concentrated. The resultant residuewas purified by column chromatography on silica gel (EtOAc/Et₃N=9/1) togive Intermediate 240-II (0.73 g) in a 80% yield.

Intermediate 240-II (0.5 g) dissolved in THF (10 mL) was added to 0.5 Mof an LiOH aqueous solution (10 mL). The mixture was stirred at roomtemperature for 2 hours. It was then acidified with 2M HCl to obtain acrude product, which was purified by column chromatography on silica gel(EtOAc/MeOH=20/1) to afford Intermediate 240-III (170 mg) in a 35%yield.

Intermediate 240-III (170 mg) was treated with 20% TFA/CH₂Cl₂ (5 mL) atroom temperature for 12 hours and then concentrated. The resultantresidue was purified by column chromatography on silica gel (21% NH₃(aq)/MeOH=1/19) to afford Compound 240 (100 mg) in a 85% yield. Compound240 was then treated with 1 M HCl (3 mL) in CH₂Cl₂ (2 mL) for 0.5 hour.After the solvents were removed, the residue was treated with ether andfiltered to give hydrochloride salt of Compound 240.

CI-MS (M⁺+1): 545.4.

EXAMPLE 241 Preparation of Compound 241

Compound 241 was prepared in a manner similar to that used to preparecompound 240.

CI-MS (M⁺+1): 558.5.

EXAMPLE 242 Preparation of Compound 242

Compound 242 was prepared in a manner similar to that used to preparecompound 240.

CI-MS (M⁺+1): 560.4.

EXAMPLE 243 Preparation of Compound 243

Compound 243 was prepared in a manner similar to that used to preparecompound 240.

CI-MS (M⁺+1): 576.4.

EXAMPLE 244 Preparation of Compound 244

Compound 244 was prepared in a manner similar to that used to preparecompound 240.

CI-MS (M⁺+1): 559.4.

EXAMPLE 245 Preparation of Compound 245

Compound 245 was prepared in a manner similar to that used to preparecompound 240.

CI-MS (M⁺+1): 531.4.

EXAMPLE 246 Preparation of Compound 246

Compound 246 was prepared in a manner similar to that used to preparecompound 240.

CI-MS (M⁺+1): 517.4.

EXAMPLE 247 Preparation of Compound 247

Compound 247 was prepared in a manner similar to that used to preparecompound 240.

CI-MS (M⁺+1): 515.4.

EXAMPLE 248 Preparation of Compound 248

Compound 248 was prepared in a manner similar to that used to preparecompound 240.

CI-MS (M⁺+1): 531.4.

EXAMPLE 249 Preparation of Compound 249

Intermediate 222-III (4.56 g) was added to a solution of2-aminoethylaniline (2.92 g) in MeOH (300 mL). The mixture was stirredat 60° C. for 8 hours. NaBH₄ (0.68 g) was then added at 0° C. for 0.5hour and the mixture was concentrated by removing the solvent. Anaqueous solution of NH₄Cl (10%, 10 mL) was added to the resultantresidue. The mixture was extracted with CH₂Cl₂, dried over anhydrousMgSO₄, filtered, and concentrated. The residue thus obtained waspurified by column chromatography on silica gel (EtOAc/MeOH=1/1) toafford Intermediate 249-I (4.2 g) in a 63% yield.

A solution of Intermediate 249-I (4.2 g) and Boc₂O (2.8 g) in CH₂Cl₂(250 mL) was added to Et₃N (1.4 mL) at 25° C. overnight. The solutionwas then concentrated and the resultant residue was purified by columnchromatography on silica gel (EtOAc/Hexane=1/5) to give Intermediate249-II (4 g) in a 75% yield.

Intermediate 249-II (4.0 g) in MeOH (20 mL) was hydrogenated in thepresence of 10% Pd/C (800 mg) and 5% Rh/C (400 mg) at 50 psi at roomtemperature for 18 hours. The mixture was then filtered andconcentrated. The residue was purified by column chromatography onsilica gel (using EtOAc/MeOH as an eluant) to afford Intermediate249-III (2.8 g) in a 69% yield.

Intermediate 249-III (900 mg) and Et₃N (0.4 mL) in 1-pentanol (5 mL) wasreacted with 2,4-dichloro-6-aminopyrimidine (365 mg) at 120° C. for 24hours. The solvent was then removed and the resultant residue waspurified by column chromatography on silica gel (EtOAc/Hexane=1/1) toafford Intermediate 249-IV (842 mg) in a 74% yield.

N¹-Morpholine-N¹-piperazine ethane (300 mg) was added to Intermediate249-IV (300 mg) in 1-pentanol (1 mL). The mixture was stirred at 120° C.for 18 hours. The solution was concentrated to give a residue, which wasthen coated with SiO₂ and purified by column chromatography on silicagel (EtOAc/MeOH=7/3) to afford Intermediate 249-V (243 mg) in a 64%yield.

A solution of 20% TFA/CH₂Cl₂ (5 mL) was added to a solution ofIntermediate 249-V (243 mg) in CH₂Cl₂ (2 mL). The reaction mixture wasstirred for 8 hours at room temperature and concentrated by removing thesolvent. The resultant residue was purified by column chromatography onsilica gel (21% NH₃ (aq)/MeOH=1/19) to afford Compound 249 (151 mg) in a84% yield. Compound 249 was t\hen treated with 1 M HCl (4 mL) in CH₂Cl₂(2 mL) for 0.5 hour. After the solvents were removed, the residue wastreated with either and filtered to give hydrochloride salt of Compound249.

CI-MS (M⁺+1): 572.5.

EXAMPLE 250 Preparation of Compound 250

Compound 250 was prepared in a manner similar to that used to preparecompound 249.

CI-MS (M⁺+1): 531.4.

EXAMPLE 251 Preparation of Compound 251

Compound 251 was prepared in a manner similar to that used to preparecompound 249.

CI-MS (M⁺+1): 547.4.

EXAMPLE 252 Preparation of Compound 252

A solution of 2-aminoethylaniline (5.0 g) and Boc₂O (6.8 g) in CH₂Cl₂(200 mL) was stirred at 25° C. overnight. The solution was thenconcentrated and the resultant residue was purified by columnchromatography on silica gel (EtOAc/Hexane=1:1 as eluant) to giveIntermediate 252-I (6.8 g) in a 83% yield.

222-III (7.3 g) prepared from Example 222 was added to a solution ofIntermediate 252-I (6.8 g) in CH₂Cl₂ (250 mL). The mixture was stirredat 25° C. for 1.5 hour. NaBH(OAc)₃ (6.0 g) and a small amount of MeOHwere added at 0° C. The mixture was stirred at room temperatureovernight. After the solution was concentrated, a saturated solution ofNaHCO₃ (250 mL) was added. The mixture was extracted with EtOAc, driedover anhydrous MgSO₄, filtered, and cocnetrated to afford crudeIntermediate 252-II (6.0 g).

Crude Intermediate 252-II (3.0 g) in MeOH (15 mL) was hydrogenated inthe presence of 5% Rh/C (300 mg) and 10% Pd/C (300 mg) at 50 psi at roomtemperature for 72 hours. The mixture was then filtered andconcentrated. The resultant residue was purified by columnchromatography on silica gel (EtOAc/MeOH=1:1) to afford Intermediate252-III (2.6 g) in a 87% yield.

A solution of intermediate 252-III (1.5 g) treated with 1M HCl in ether(52 mL) and MeOH (10 mL) was stirred at room temperature for 8 hours.After dditional ether was added, the solution was filtered. The solidthus obtained was dried under vacuum. K₂CO₃ was added to a suspension ofthe solid in CH₃CN at room temperature for 10 minutes. After water wasadded, the reaction mixture was stirred at room temperature for 2 hours,filtered, dried over anhydrous MgSO₄, and concentrated to afford crudeIntermediate 252-IV (1.5 g).

Intermediate 252-IV (1.5 g) and Et₃N (0.5 mL) in 1-pentanol (14 mL) wasallowed to react with 2,4-dichloro-6-aminopyrimidine (1.0 g) at 120° C.overnight. The solvent was then removed to afford crude Intermediate252-V (2.0 g).

A solution of Intermediate 252-V (2.0 g) and Boc₂O (2.1 g) in CH₂Cl₂(250 mL) was added to Et₃N (1.0 mL) at 25° C. overnight. The solutionwas then concentrated and the resultant residue was purified by columnchromatography on silica gel (EtOAc/Hexane=1:1) to give Intermediate252-VI (1.7 g) in a 56% yield.

N¹-Morpholine-N¹-piperazine ethane (300 mg) was added to Intermediate252-VI (300 mg) in 1-pentanol (1 mL). The mixture was stirred at 120° C.overnight and then concentrated. The residue thus obtained was coatedwith SiO₂ and purified by column chromatography on silica gel(EtOAc/MeOH=1/1) to afford Intermediate 252-VII (260 mg) in a 70% yield.

A solution of 20% TFA/CH₂Cl₂ (5 mL) was added to a solution ofIntermediate 252-VII (260 mg) in CH₂Cl₂ (2 mL). The reaction mixture wasstirred for 8 hours at room temperature and concentrated by removing thesolvent. The resultant residue was purified by column chromatography onsilica gel (21% NH₃ (aq)/MeOH=1/19) to afford Compound 252 (175 mg) in a91% yield. Compound 252 was then treated with 1 M HCl (4 mL) in CH₂Cl₂(2 mL) for 0.5 hours. After the solvents were removed, the residue wastreated with ether and filtered to give hydrochloride salt of Compound252.

CI-MS (M⁺+1): 572.5.

EXAMPLE 253 Preparation of Compound 253

Compound 253 was prepared in a manner similar to that used to preparecompound 252.

CI-MS (M⁺+1): 531.4.

EXAMPLE 254 Preparation of Compound 254

Compound 254 was prepared in a manner similar to that used to preparecompound 252.

CI-MS (M⁺+1): 547.4.

EXAMPLE 255 Preparation of Compound 255

A solution of Intermediate 252-VI (1.0 g) and piperazine (0.42 g) in1-pentanol (8 mL) was stirred at 120° C. overnight. After the solutionwas concentrated, the residue was treated with water and extracted withCH₂Cl₂. The organic layer was separated and concentrated. The residuethus obtained was purified by column chromatography on silica gel(EtOAc/MeOH=2/1) to afford Intermediate 255-I (0.9 g) in a 84% yield.

To a solution of Intermediate 255-I (0.4 g) in CH₃CN (6 mL) were addedethyl bromoacetate (100 mg) and K₂CO₃ (400 mg). The mixture was stirredat 60° C. for 3 hours. After the solution was filtered and concentrated,the residue was purified by column chromatography on silica gel(EtOAc/MeOH=4/1) to afford Intermediate 255-II (0.17 g) in a 38% yield.

Intermediate 255-II (0.17 g) dissolved in THF (2 mL) was added to 0.5 Mof an LiOH aqueous solution (2 mL). The mixture was stirred at roomtemperature for 15 hours. It was acidified with 2.5 M HCl (PH=9) andfiltered to obtain yellow solid. The yellow solid was purified by columnchromatography on silica gel (EtOAc/MeOH=1/5) to afford Intermediate255-III (0.1 g) in a 61% yield.

20% TFA/CH₂Cl₂ (3 mL) was added to a solution of Intermediate 255-III(100 mg) in CH₂Cl₂ (2 mL). The solution was stirred at room temperaturefor 2 hours and then concentrated. The residue in acetone (3 mL) wasadded to HCl (4 M in dioxane, 1 mL) at room temperature and the mixturewas stirred for 30 minutes. After the solvents were removed, the residuewas treated with ether and filtered to give hydrochloride salt ofCompound 255 (62 mg).

CI-MS (M⁺+1): 517.4.

EXAMPLE 256 Preparation of Compound 256

Compound 256 was prepared in a manner similar to that used to preparecompound 255.

CI-MS (M⁺+1): 531.7.

EXAMPLE 257 Preparation of Compound 257

KOH (14 g) and Boc₂O (33.3 g) were added to a solution oftrans-4-(Aminomethyl)cyclohexane-carboxylic acid (20 g) in dioxane (112mL) at 0° C. The reaction was stirred at 25° C. overnight. The solutionwas concentrated to half of the original volume under vacuum, acidifiedwith 2.5 N HCl (PH=3), and extracted with EtOAc. The combined organiclayer was washed with brine, dried over anhydrous MgSO₄, filtered, andconcentrated to give a white solid Intermediate 257-I (31.9 g).

To a suspension of the above solid in toluene (150 mL) were addedphosphorazidic acid diphenyl ester (32.4 g) and Et₃N (11.9 g) at 25° C.for 1.0 hour. The reaction mixture was warmed to 80° C. for 3.0 hoursand then cooled to 25° C. After benzyl alcohol (20 g) was added, thereaction mixture was stirred at 80° C. for another 3.0 hours and thenwarmed to 120° C. overnight. It was then concentrated and dissolvedagain in EtOAc and H₂O. The organic layer was then collected. Theaqueous layer was extracted with EtOAc. The combined organic layer waswashed with 2.5 N HCl, saturated aqueous NaHCO₃ and brine, dried overanhydrous MgSO₄, filtered, and concentrated. The residue thus obtainedwas purified by column chromatography on silica gel (EtOAc/Hexane=1/2)to give Intermediate 257-II (35 g) in a 79% yield.

To a suspension of Intermediate 257-II (1.9 g) in MeOH (10 mL) was added10% Pd/C (190 mg). The mixture was stirred at ambient temperature underhydrogen atmosphere for 4.0 hours, filtered, and concentrated. Theresidue thus obtained was purified by column chromatography on silicagel (using EtOAc and MeOH as an eluant) to give Intermediate 257-III(750 mg) in a 60% yield.

222-III (1,198 mg) prepared from Example 222 was added to a solution ofIntermediate 257-III (750 mg) in CH₂Cl₂ (30 mL). The mixture was stirredat 25° C. for 2 hours. NaBH(OAc)₃ (1,046 mg) was then added at 25° C.for 12 hours. After the solution was concentrated, a saturated aqueousNaHCO₃ solution was added to the resultant residue. The mixture wasextracted with CH₂Cl₂. The organic layer was collected and concentrated.The residue thus obtained was purified by column chromatography onsilica gel (using EtOAc and MeOH as an eluant) to afford Intermediate257-IV (1,200 mg) in a 78% yield.

A solution of Intermediate 257-IV (5.2 g) treated with 4 N HCl/dioxane(39 mL) in MeOH (52 mL) was stirred at room temperature for 8 hours.After ether (104 mL) was added, the solution was filtered. The solidthus obtained was dried under vacuum. K₂CO₃ (21 g) was added to asuspension of this solid in CH₃CN (230 mL) at room temperature for 10minutes. After water (9 mL) was added, the reaction mixture was stirredat room temperature for 2 hours. The mixture was then filtered, driedover anhydrous MgSO₄, and concentrated to afford crude Intermediate257-V (2.8 g).

Crude Intermediate 257-V (2.8 g) and Et₃N (1.3 mL) in 1-pentanol (11.3mL) was allowed to react with 2,4-dichloro-6-aminopyrimidine (1,633 mg)at 100° C. for 12 hours. The solvent was then removed and the residuewas purified by column chromatography on silica gel (21% NH₃(aq)/MeOH=1/19) to afford Intermediate 257-VI (3.3 g) in a 75% yield.

A solution of Intermediate 252-VI (3.3 g) and Boc₂O (4.189 g) in CH₂Cl₂(60 mL) was added to Et₃N (1.0 mL) at 25° C. overnight. The solution wasthen concentrated and the resultant residue was purified by columnchromatography on silica gel (using EtOAc and Hexane as an eluant) togive Intermediate 257-VII (3.2 g) in a 64% yield.

Intermediate 257-VII (2.6 g) and piperazine (1.127 g) in 1-pentanol (5.2mL) was added to Et₃N (0.5 mL) at 120° C. for 18 hours. After thesolution was concentrated, the residue was treated with water andextracted with CH₂Cl₂. The organic layer was collected and concentrated.The residue thus obtained was purified by column chromatography onsilica gel (using EtOAc/MeOH to 21% NH₃ (aq)/MeOH as an eluant) toafford Intermediate 257-VIII (1.8 g) in a 64% yield.

To a solution of Intermediate 257-VIII (200 mg) in CH₃CN (20 mL) wereadded ethyl bromoacetate (52 mg) and K₂CO₃ (128 mg). The mixture wasstirred at 60° C. for 2 hours. The solution was filtered andconcentrated. The residue was purified by column chromatography onsilica gel (using EtOAc and MeOH as an eluant) to afford Intermediate257-IX (140 mg) in a 62% yield.

0.5 M of a LiOH aqueous solution (10 mL) was added to Intermediate257-IX (500 mg) dissolved in THF (10 mL). The mixture was stirred atroom temperature for 15 hours. It was then acidified with 2.5 M HCl(pH=9) and filtered to obtain a yellow solid. The yellow solid waspurified by column chromatography on silica gel (using EtOAc/MeOH to 21%NH₃ (aq)/MeOH as an eluant) to afford Intermediate 257-X (337 mg) in a70% yield.

20% TFA/CH₂Cl₂ (10 mL) was added to a solution of Intermediate 257-X(400 mg) in CH₂Cl₂ (8 mL). The solution was stirred at room temperaturefor 2 hours and then concentrated. To the residue in acetone (7 mL) wasadded HCl (4 M in dioxane, 1.3 mL) at room temperature for 30 minutes.After the solvents were removed, the residue was treated with ether andfiltered to give hydrochloride salt of Compound 257 (257 mg).

CI-MS (M⁺+1): 503.4.

EXAMPLE 258 Preparation of Compound 258

Compound 258 was prepared in a manner similar to that used to preparecompound 257.

CI-MS (M⁺+1): 531.4.

EXAMPLE 259 Preparation of Compound 259

Compound 259 was prepared in a manner similar to that used to preparecompound 257.

CI-MS (M⁺+1): 517.4.

EXAMPLE 260 Preparation of Compound 260

A solution of Intermediate 257-II (35 g) treated with 4 N HCl/dioxane(210 mL) in MeOH (350 mL) was stirred at room temperature overnight.After ether (700 mL) was added, the solution was filtered. The solidthus obtained was dried under vacuum. K₂CO₃ was added to a suspension ofthis solid in CH₃CN and iso-propanol at room temperature for 10 minutes.After water was added, the reaction mixture was stirred at roomtemperature for 2 hours, filtered, dried over anhydrous MgSO₄, andconcentrated. The resultant residue was purified by columnchromatography on silica gel (using CH₂Cl₂ and MeOH as an eluant) togive Intermediate 260-I (19 g) in a 76% yield.

Intermediate 222-III (21 g) prepared from Example 222 was added to asolution of Intermediate 260-I (19 g) in CH₂Cl₂ (570 mL). The mixturewas stirred at 25° C. for 2 hours. NaBH(OAc)₃ (23 g) was then added at25° C. overnight. After the solution was concentrated, a saturatedaqueous NaHCO₃ solution was added to the resultant residue. The mixturewas then extracted with CH₂Cl₂. The solution was concentrated and theresidue was purified by column chromatography on silica gel (using EtOAcand MeOH as an eluant) to afford Intermediate 260-II (23.9 g) in a 66%yield.

A solution of Intermediate 260-II (23.9 g) and Boc₂O (11.4 g) in CH₂Cl₂(200 mL) was added to Et₃N (5.8 mL) at 25° C. for overnight. Thesolution was then concentrated and the resultant residue was purified bycolumn chromatography on silica gel (using EtOAc and Hexane as aneluant) to give Intermediate 260-III (22 g) in a 77% yield.

10% Pd/C (2.2 g) was added to a suspension of Intermediate 260-III (22g) in MeOH (44 mL). The mixture was stirred at ambient temperature underhydrogen atmosphere overnight, filtered, and concentrated. The residuethus obtained was purified by column chromatography on silica gel (usingEtOAc and MeOH as an eluant) to afford Intermediate 260-IV (16.5 g) in a97% yield.

Intermediate 260-IV (16.5 g) and Et₃N (4.4 mL) in 1-pentanol (75 mL) wasallowed to react with 2,4-dichloro-6-aminopyrimidine (21 g) at 120° C.overnight. The solvent was then removed and the residue was purified bycolumn chromatography on silica gel (using EtOAc and hexane as aneluant) to afford Intermediate 260-V (16.2 g) in a 77% yield.

Intermediate 260-V (16.2 g) and piperazine (11.7 g) in 1-pentanol (32mL) was added to Et₃N (3.3 mL) at 120° C. overnight. After the solutionwas concentrated, the residue was treated with water and extracted withCH₂Cl₂. The organic layer was collected and concentrated. The residuethus obtained was purified by column chromatography on silica gel (usingEtOAc/MeOH to 21% NH₃ (aq)/MeOH as an eluant) to afford Intermediate260-VI (13.2 g) in a 75% yield.

Methylacrylate (532 mg) was added to a solution of Intermediate 260-VI(4 g) in MeOH (200 mL) at 25° C. for 5 hours. The solution was thenconcentrated and the resultant residue was purification by columnchromatography on silica gel (using EtOAc and MeOH as an eluant) toafford Intermediate 260-VII (3 g) in a 66% yield.

Intermediate 260-VII (3 g) dissolved in THF (60 mL) was added 0.5 M of aLiOH aqueous solution (60 mL). The mixture was stirred at roomtemperature for 15 hours. It was then acidified with 2.5 M HCl (pH=8)and filtered to obtain a yellow solid. The yellow solid was purified bycolumn chromatography on silica gel (using CH₂Cl₂ and MeOH as an eluant)to afford Intermediate 260-VIII (1.4 g) in a 48% yield

20% TFA/CH₂Cl₂ (34 mL) was added to a solution of Intermediate 260-VIII(1.4 g) in CH₂Cl₂ (17 mL). The solution was stirred at room temperaturefor 5 hours and then concentrated. To the residue in acetone (20 mL) wasadded HCl (4 M in dioxane, 4 mL) at room temperature for 30 minutes.After the solvents were removed, the residue was treated with ether and(20 mL) and filtered to give hydrochloride salt of Compound 260 (1.4 g).

CI-MS (M⁺+1): 517.4.

EXAMPLE 261 Preparation of Compound 261

Compound 261 was prepared in a manner similar to that used to preparecompound 260.

CI-MS (M⁺+1): 531.4.

EXAMPLE 262 Preparation of Compound 262

Compound 262 was prepared in a manner similar to that used to preparecompound 260.

CI-MS (M⁺+1): 503.4.

EXAMPLE 263 Preparation of Compound 263

Compound 263 was prepared in a manner similar to that used to preparecompound 260.

CI-MS (M⁺+1): 545.4.

EXAMPLE 264 Preparation of Compound 264

Compound 264 was prepared in a manner similar to that used to preparecompound 260.

CI-MS (M⁺+1): 545.4.

EXAMPLE 265 Preparation of Compound 265

A suspension of cis-4-amino-cyclohexanecarboxylic acid (20 g) in MeOH(300 mL) was cooled to 0° C. Thionyl chloride (12.2 mL) was addeddropwise to the suspension. The mixture was stirred at room temperatureovernight and concentrated. To the he residue in CH₃CN was added K₂CO₃at room temperature for 10 minutes. After water was added, the mixturewas stirred at room temperature for 2 hours, filtered, dried overanhydrous MgSO₄, concentrated to give a white solid Intermediate 265-I(25 g).

Boc₂O (31.8 g) was added to a suspension of the crude Intermediate 265-I(25 g) in CH₂Cl₂ (300 mL) at 0° C. for 0.5 hour. The reaction mixturewas stirred at 25° C. overnight and poured into water. The aqueous layerwas extracted with CH₂Cl₂. The combined organic layer was collected,dried over anhydrous MgSO₄, filtered, and concentrated to give crudeIntermediate 265-II (38 g).

A solution of crude Intermediate 265-II (38 g) in Et₂O (100 mL) wasadded to a suspension of LiAlH₄ (6.7 g) in Et₂O (400 mL) below 0° C. Thereaction mixture was stirred at room temperature overnight. The mixturewas quenched with Na₂SO₄.10H₂O, and filtered through a pad of celite.The filtrate was dried over anhydrous MgSO₄, filtered, dried underreduced pressure. The residue thus obtained was purified by columnchromatography on silica gel (using CH₂Cl₂ and MeOH as an eluant) toafford Intermediate 265-III (28 g) in a 90% yield.

To a solution of Intermediate 265-III (28 g) in CH₂Cl₂ (300 mL) wereadded Et₃N (30 mL), DMPA (0.7 g), and p-toluenesulfonyl chloride (25.8g). The reaction mixture was stirred at 25° C. overnight. The resultingsolution was concentrated and the residue was re-dissolved in EtOAc. Thesolution was then washed with water and extracted with EtOAc. Theorganic layer was collected, dried over anhydrous MgSO₄, concentrated togive Intermediate 265-IV (50 g).

NaN₃ (24 g) was added to a solution of Intermediate 265-IV (50 g) in DMF(300 mL). The resulting mixture was stirred at 60° C. overnight,filtered, and concentrated. The residue in CH₂Cl₂ was washed with asaturated aq. NaHCO₃ solution. The organic solution collected, dried,and concentrated to give a residue. The residue was purified by columnchromatography on silica gel (using EtOAc as an eluant) to affordIntermediate 265-V (30 g) in a 97% yield.

To a suspension of LiAlH₄ (5.4 g) in Et₂O (400 mL) was added a solutionof crude Intermediate 265-V (30 g) in Et₂O (100 mL) below 0° C. Thereaction mixture was stirred at room temperature overnight. The reactionwas quenched with Na₂SO₄.10H₂O, filtered through a pad of celite. Thefiltrate was dried over anhydrous MgSO₄, filtered, and dried underreduced pressure. The residue thus obtained was purified by columnchromatography on silica gel (MeOH as an eluant) to afford Intermediate265-VI (24.5 g) in a 90% yield.

A solution of Intermediate 265-VI (24.5 g) and Et₃N (13 mL) in1-pentanol (75 mL) was reacted with 2,4-dichloro-6-aminopyrimidine (19.6g) at 120° C. overnight. The reaction mixture was stirred at 150° C. for3 hours, filtered, and dried under reduced pressure. The residue thusobtained was purified by column chromatography on silica gel (EtOAc asan eluant) to afford Intermediate 265-VII (26.2 g) in a 68% yield.

A solution of Intermediate 265-VII (26.2 g) treated with 4 N HCl/dioxane(160 mL) in MeOH (200 mL) was stirred at room temperature overnight.After ether was added, the solution was filtered. The solid thusobtained was dried by vacuum. To a suspension of the above solid inCH₃CN and iso-propanol was added K₂CO₃ at room temperature for 10minutes. After water was added at room temperature for 2 hours, thereaction mixture was filtered, dried over anhydrous MgSO₄, filtered, andconcentrated and to give Intermediate 265-VIII (15 g).

To a solution of Intermediate 265-VIII (15 g) in CH₂Cl₂ (500 mL) wasadded Intermediate 222-III (18.6 g). The mixture was stirred at 25° C.for 2 hours. NaBH(OAc)₃ (11.7 g) was then added at 25° C. and themixture was stirred overnight. The solution was then concentrated and asaturated aqueous NaHCO₃ solution was added. The mixture was extractedwith CH₂Cl₂. The organic layer was collected, dried over anhydrousMgSO₄, concentrated. The residue thus obtained was purified by columnchromatography on silica gel (MeOH as an eluant) to afford Intermediate265-IX (14.1 g) in a 39% yield.

Et₃N (2.2 mL) was added to a solution of Intermediate 265-IX (14.1 g)and Boc₂O (6.6 g) in CH₂Cl₂ (150 mL) at 25° C. The solution was stirredovernight and then concentrated. The resultant residue was purified bycolumn chromatography on silica gel (EtOAc as an eluant) to giveIntermediate 265-X (12 g) in a 71% yield.

Et₃N (2.4 mL) was added to a mixture of Intermediate 265-X (12 g) andpiperazine (5.1 g) in 1-pentanol (24 mL) 120° C. The solution wasstirred overnight and then concentrated. The residue was treated withwater and extracted with CH₂Cl₂. The organic layer was collected, driedover anhydrous MgSO₄. The residue thus obtained was purified by columnchromatography on silica gel (MeOH as eluant) to afford Intermediate265-XI (9.6 g) in a 74% yield.

To a solution of Intermediate 265-XI (500 mg) in CH₃CN (50 mL) wereadded ethyl bromoacetate (127 mg) and K₂CO₃ (314 mg). The mixture wasstirred at 60° C. for 2 hours. The solution was filtered andconcentrated. The residue thus obtained was purified by columnchromatography on silica gel (EtOAc as an eluant) to afford Intermediate265-XII (230 mg) in a 41% yield.

0.5 M LiOH (5 mL) was added to a solution of Intermediate 265-XII (230mg) in THF (10 mL). The mixture was stirred at room temperature for 15hours. It was then acidified with 2.5 M HCl (pH=8) and filtered toobtain a yellow solid, which was purified by column chromatography onsilica gel (MeOH as an eluant) to afford Intermediate 265-XIII (150 mg)in a 68% yield.

To a solution of Intermediate 265-XIII (150 mg) in CH₂Cl₂ (2 mL) wasadded 20% TFA/CH₂Cl₂ (3 mL). The solution was stirred at roomtemperature for 5 hours and then concentrated. HCl (4 M in dioxane, 2mL) was added in the residue in acetone at room temperature for 30minutes. After solvents were removed, the residue was treated with etherand filtered to give hydrochloride salt of compound 265 (94 mg).

CI-MS (M⁺+1): 517.4.

EXAMPLE 266 Preparation of Compound 266

Compound 266 was prepared in a manner similar to that used to preparecompound 265 (see example 264).

CI-MS (M⁺+1): 531.4.

EXAMPLE 267 Preparation of Compound 267

A suspension of cis-4-amino-cyclohexanecarboxylic acid (30 g) in MeOH(500 mL) was cooled to 0° C. Thionyl chloride (30.5 mL) was addeddropwise. The mixture was stirred at room temperature overnight andconcentrated to give a white solid Intermediate 267-I (32.6 g).

To a suspension of Intermediate 267-I (32.6 g) obtained above in CH₂Cl₂(500 mL) were added Et₃N (18 mL) and Boc₂O (50 g) sequentially. Thereaction mixture was stirred at 25° C. overnight and poured into water.The aqueous layer was extracted with CH₂Cl₂. The combined organic layerwas dried over anhydrous MgSO₄, filtered, concentrated to give crudeIntermediate 267-II (53.4 g).

A solution of crude Intermediate 267-II (53.4 g) in Et₂O (100 mL) wasadded to a suspension cooled at 0° C. of LiAlH₄ (11 g) in Et₂O (500 mL)below 0° C. The reaction mixture was stirred at room temperatureovernight. The resulting solution was cooled in ice-bath, quenched withcold water, filtered through a pad of celite. The filtrate was driedover anhydrous MgSO₄, filtered, washed with hexane, and dried underreduced pressure to give crude Intermediate 267-III (43.21 g).

Et₃N (32 mL), DMPA (4.6 g) and p-toluenesulfonyl chloride (40 g) wereadded to a solution of Intermediate 267-III (43.21 g) in CH₂Cl₂ (400mL). The reaction mixture was stirred at 25° C. overnight. The resultingsolution was concentrated and the residue was dissolved in EtOAc. Thesolution was washed with water and extracted with EtOAc. The organiclayer was dried over anhydrous MgSO₄, and concentrated to give aresidue. The residue was purified by column chromatography on silica gel(EtOAc/Hexane=1/4) to afford Intermediate 267-IV (57.34 g) in a 71%yield.

NaN₃ (29 g) was added to a solution of Intermediate 267-IV (57.34 g) inDMF (200 mL). The resulting mixture was stirred at 40° C. overnight,filtered, and concentrated. The residue was dissolved in CH₂Cl₂ and waswashed with a saturated aqueous NaHCO₃ solution. The solution wasconcentrated to give a residue, which was purified by columnchromatography on silica gel (EtOAc/Hexane=1/6) to afford Intermediate267-V (30.48 g) in a 80% yield.

PPh₃ (12.9 g) and H₂O (0.9 mL) were added to a solution of Intermediate267-V (11.37 g) in THF (200 mL). After the solution was stirred at 25°C. overnight, the solution was concentrated to give a residue, which waspurified by column chromatography on silica gel (EtOAc/MeOH=15/1) toafford Intermediate 267-VI (9.44 g) in a 93% yield.

A solution of Intermediate 267-VI (9.44 g) and Et₃N (4 mL) in 1-pentanol(40 mL) was reacted with 2,4-dichloro-6-aminopyrimidine (7.5 g). Thesolution was stirred at 120° C. overnight. The solvent was removed andthe residue thus obtained was purified by column chromatography onsilica gel (EtOAc/MeOH=1/2) to afford Intermediate 267-VII (10.5 g) in a71% yield.

A solution of Intermediate 267-VII (2.0 g) treated with 4 N HCl/dioxane(10 mL) in MeOH (20 mL) was stirred at room temperature overnight. Afterether was added, the solution was filtered. The solid was dried undervacuum. To a suspension of the above solid in CH₃CN and iso-propanol wasadded K₂CO₃ at room temperature and was stirred for 10 minutes. Afterwater was added to the reaction mixture at room temperature, it wasstirred for another 2 hours. The mixture was then filtered, dried overanhydrous MgSO₄, filtered, and concentrated. The resultant residue waspurified by column chromatography on silica gel (CH₂Cl₂ and MeOH aseluant) to give Intermediate 267-VIII (1.1 g) in a 77% yield.

Intermediate 222-III (1.58 g) was added to a solution of Intermediate267-VIII (1.10 g) in CH₂Cl₂ (40 mL). The mixture was stirred at 25° C.for 2 hours. NaBH(OAc)₃ (907 mg) was then added at 25° C. and themixture was stirred overnight. The mixture was then concentrated and asaturated aqueous NaHCO₃ solution was added to the resultant residue.The mixture was extracted with CH₂Cl₂. The organic layer was collectedand concentrated. The residue thus obtained was purified by columnchromatography on silica gel (EtOAc and MeOH as eluant) to affordIntermediate 267-IX (1.30 g) in a 61% yield.

Et₃N (0.3 mL) was added to a solution of Intermediate 267-IX (1.30 g)and Boc₂O (0.63 g) in CH₂Cl₂ (150 mL) at 25° C. The solution was stirredovernight and then concentrated. The resultant residue was purified bycolumn chromatography on silica gel (EtOAc/Hexane=1/1) to giveIntermediate 267-X (1.30 g) in a 83% yield.

Et₃N (0.2 mL) was added to a solution of Intermediate 267-X (800 mg) andpiperazine (347 mg) in 1-pentanol (2 mL) at 120° C. The solution wasstirred overnight and then concentrated. The residue was treated withwater and extracted with CH₂Cl₂. The organic layer was collected andconcentrated. The residue thus obtained was purified by columnchromatography on silica gel (EtOAc/MeOH=1/1) to afford Intermediate267-XI (700 mg) in a 81% yield.

Methylacrylate (93 mg) was added to a solution of Intermediate 267-XI(700 mg) in MeOH (7 mL) at 30° C. The solution was stirred for 5 hoursand then concentrated. The residue was purified by silica gel (EtOAc andMeOH as eluant) to afford Intermediate 267-XII (460 mg) in a 58% yield.

0.5 M LiOH (9.2 mL) was added to a solution of Intermediate 267-XII (460mg) in THF (5 mL). The mixture was stirred at room temperature for 15hours. It was then acidified with 2.5 M HCl (pH=8) and filtered toobtain a yellow solid. The solid was purified by column chromatographyon silica gel (CH₂Cl₂ and MeOH as eluant) to afford Intermediate267-XIII (266 mg) in a 59% yield

20% TFA/CH₂Cl₂ (5 mL) was added to a solution of Intermediate 267-XIII(266 mg) in CH₂Cl₂ (3 mL). The solution was stirred at room temperaturefor 5 hours. The solution was concentrated. HCl (4 M in dioxane, 3 mL)was added to the residue in acetone. The mixture was stirred at roomtemperature for 30 minutes. After solvents were removed, the residue wastreated with ether and filtered to give the hydrochloride salt ofCompound 267 (153 mg).

CI-MS (M⁺+1): 517.4.

EXAMPLE 268 Preparation of Compound 268

A solution of methyl cis-4-(amino)cyclohexylcarboxylate (267-I, 5.0 g)and PhCH₂OCOCl (6.5 g) in CH₂Cl₂ (64 mL) was stirred at 0° C. for 1hour. The solution was allowed to warm-up to room temperature andstirred for another 12 hours. It was then concentrated and the residuewas purified by column chromatography on silica gel (EtOAc/Hexane=1/4)to afford Intermediate 268-I (5.56 g) in a 60% yield.

DIBAL (1.0 M in Hexane, 34 mL) was added to a stirred solution ofIntermediate 268-I (5.0 g) in dry toluene (170 mL) at −70˜−78° C. underN₂ (g). The reaction mixture was stirred for 2 hours at thistemperature. 5% HCl (aq) (34 mL) was then added to the solution at−60˜−70° C. and the mixture was stirred for another 0.5 hour after thereaction temperature was increased to 25° C. The aqueous layer wasextracted with CH₂Cl₂ twice. The organic layers were combined, driedwith anhydrous MgSO₄, and concentrated by removing the solvent undervacuum to afford curd 268-II (3.14 g).

Intermediate 268-II (3.0 g) was then reacted at room temperature withcyclohexylaminopropylamine (1.8 g) in MeOH (30 mL). The mixture wasstirred at 60° C. for 12 hours. NaBH₄ (0.43 g) was added at 0° C. Afterbeing stirred for 0.5 hour, an aqueous solution of NH₄Cl (10%, 20 mL)was added and the mixture was extracted with CH₂Cl₂. The organic layerwas collected, dried over anhydrous MgSO₄, filtered, and concentrated toafford a residue. The residue was purified by column chromatography onsilica gel (EtOAc/Hexane=4/1) to afford Intermediate 268-III (2.54 g) ina 55% yield.

A solution of Intermediate 268-III (2.5 g) and Boc₂O (3.0 g) in CH₂Cl₂(130 mL) was stirred at 25° C. overnight. The solution was thenconcentrated and the resultant residue was purified by columnchromatography on silica gel (EtOAc/Hexane=1/9) to give Intermediate268-IV (3.2 g) in a 85% yield.

10% Pd/C (570 mg) was added to a suspension of Intermediate 268-IV (3.2g) in EtOH (18 mL). The mixture was stirred at ambient temperature underhydrogen atmosphere for 2.0 hours, filtered, and concentrated to givecrude Intermediate 268-V (2.4 g).

A solution of crude Intermediate 268-V (2.4 g) and Et₃N (0.85 mL) in1-pentanol (17 mL) was reacted with 2,4-dichloro-6-aminopyrimidine (1.0g) at 120° C. for 15 hours. The solvent was removed and the residue waspurified by column chromatography on silica gel (EtOAc/Hexane=3/1) toafford Intermediate 268-VI (2.4 g) in a 80% yield.

Piperazine (1.0 g) was added to Intermediate 268-VI (2.4 g) in1-pentanol (8 mL). The mixture was stirred at 120° C. for 15 hours. Thesolution was concentrated and the residue was treated with water andextracted with CH₂Cl₂. The organic layer was collected and concentratedto afford crude Intermediate 268-VII (2.2 g).

Methyl acrylate (0.1 mL) was added to a solution of crude Intermediate268-VII (700 mg) in MeOH (10 mL). The mixture was stirred at 30° C. for12 hours. After the mixture was concentrated, the residue was treatedwith water and extracted with CH₂Cl₂. The organic layer was collectedand concentrated to give a residue, which was purified by columnchromatography on silica gel (EtOAc/MeOH=4/1) to afford Intermediate268-VIII (530 mg) in a 67% yield.

0.5 M LiOH (7.2 mL) was added to a solution of Intermediate 268-VIII(530 mg) in THF (7.2 mL). The mixture was stirred at room temperaturefor 2 hours. It was then acidified with 2M HCl (pH=8) and filtered toobtain a yellow solid, which was purifed by column chromatography onsilica gel (EtOAc/MeOH=1/9) to afford Intermediate 268-IX (470 mg) in a90% yield.

Intermediate 268-IX (470 mg) was dissolved in CH₂Cl₂ (8 mL). TFA (2 mL)was added and the solution was stirred at room temperature overnight.The solution was then concentrated and HCl (4 M in dioxane, 1.3 mL) wasadded to the residue in acetone (7 mL) at room temperature for 30minutes. After the solvents were removed, the residue was treated withether and filtered to give a hydrochloride salt of compound 268 (390mg).

CI-MS (M⁺+1): 517.4.

EXAMPLE 269 In Vitro Assay

143 of the above-mentioned compounds were tested for their efficacy inbinding to CXCR4 receptor using a DELFIA GTP-binding kit (Wallac Oy,Turku, Finland). The DELFIA GTP-binding assay is a time-resolvedfluorometric assay based on GDP-GTP exchange on G-protein subunitsfollowed by activation of a G protein-coupled receptor by its agonists.Eu-GTP, obtained from Wallac Oy, was used in this assay to allowmonitoring of agonist-dependent activation of G-protein. Stimulation ofCXCR4 receptor by SDF-1 leads to the replacement of GDP by GTP on theα-subunit of G-protein. This GTP-Gα complex represents the activatedform of G-protein. Eu-GTP, a non-hydrolysable analog of GTP, can be usedto quantify the amount of activated G-protein. (Peltonen et al., Eur. J.Pharmacol. (1998) 355:275.)

Plasma membrane of CXCR4-expressing HEK293 cells was suspended in anassay buffer (50 mM NaCl, 100 μg/mL saponin, 3 mM MgCl₂, 3 μM GDP, 5%BSA, 50 mM HEPES, pH 7.4). An aliquot (4 μg protein) was added to eachwell of an AcroPlate (Pall Life Sciences, Ann Arbor, Mich.). After theaddition of the test compounds (10 μM in 0.1% DMSO) and stromal-derivedfactor-1 (4 nM in the assay buffer), the assay plate was incubated inthe dark at room temperature with slow shaking for 10 minutes. Eu-GTPwas added to each well and the plate was incubated again for 60 minutes.The assay was terminated by washing the plate twice with a wash solutionprovided in the assay kit. Binding of Eu-GTP was determined based on thefluorescence signal from a Victor 2 multi-label reader.

Unexpectedly, 196 of the tested compounds showed IC₅₀ values between0.003 μM and 0.1 μM; 56 of the tested compounds showed IC₅₀ valuesbetween 0.1 μM and 1 μM, and 16 of the test compounds showed IC₅₀ valuesbetween 1 μM and 5 μM.

EXAMPLE 270 Radioligand Binding Assay

Competition binding assays between test compounds human stromal-derivedfactor-1 were carried out using glass fiber filter plates (Millipore,Billerica, Mass.). The glass fiber filter plates were pre-coated with 90μl of 0.2% polyethyleneimine for 30 minutes and rinsed with 100 μl ofdistilled water for four times to reduce non-specific binding. Membranesof human CXCR4-transfected HEK293 cells (5-10 μg protein/well) in a 70μl assay buffer (50 mM HEPES, pH 7.4, 0.5% bovine serum albumin, 90 mMNaCl, 5 mM MgCl₂, 1 mM CaCl₂) were incubated with 20 μl of a testcompound solution and 10 μl of a [¹²⁵I]-SDF-1 solution (each having afinal concentration 150 pM) in U-bottom assay plates (Corning, Corning,N.Y.). After the membranes were incubated at room temperature for 120minutes, the incubation was terminated by transferring 80 μl of eachreaction mixture to each glass fiber plate well and filtered by vacuumfiltration (MultiScreen Vacuum Maniford, Millipore). Each plate waswashed 4 times with 80 μl/well of a wash buffer (20 mM HEPES, pH 7.4 and90 mM NaCl) and then air dried overnight. After 35 μl/well of a Supermixcocktail to each plate, the radioactivity retained on the plate wascounted with Trilux MicroBeta (PerkinElmer, Boston, Mass.).

The 196 compounds with IC₅₀ values between 0.003 μM and 0.1 μM inGTP-binding assay were further screened in radioligand binding assay.The results show that they exhibited inhibitory activities in the rangeof 10-1200 nM.

Other Embodiments

All of the features disclosed in this specification may be combined inany combination. Each feature disclosed in this specification may bereplaced by an alternative feature serving the same, equivalent, orsimilar purpose. Thus, unless expressly stated otherwise, each featuredisclosed is only an example of a generic series of equivalent orsimilar features.

From the above description, one skilled in the art can easily ascertainthe essential characteristics of the present invention, and withoutdeparting from the spirit and scope thereof, can make various changesand modifications of the invention to adapt it to various usages andconditions. Thus, other embodiments are also within the scope of thefollowing claims.

1. A compound of formula (I):

wherein X is —N(R_(a))— is C₁-C₁₀ alkylene or —C(O)— L₂ is deleted orC₁-C₁₀ alkylene; L₃ is C₃-C₂₀ cycloalkyl; each of R₁, R₂, and R₃,independently, is H, C₁-C₁₀ alkyl, C₃-C₂₀ cycloalkyl, C₃-C₂₀heterocycloalkyl, aryl, heteroaryl, halo, CN, OR_(c), COOR_(c),OC(O)R_(c), C(O)R_(c), C(O)NR_(c)R_(d), or NR_(c)R_(d); R₄ is H; and R₅is C₃-C₂₀ cycloalkyl, C₃-C₂₀ heterocycloalkyl, aryl, heteroaryl, orC₁-C₁₀ alkyl substituted with C₃-C₂₀ cycloalkyl, C₃-C₂₀heterocycloalkyl, or N(R_(e)R_(f)); in which each of R_(a), R_(b),R_(c), R_(d), R_(e), and R_(f), independently, is H, C₁-C₁₀ alkyl,C₃-C₂₀ cycloalkyl, C₃-C₂₀ heterocycloalkyl, aryl, heteroaryl, or —C(O)R;R being H, C₁-C₁₀ alkyl, C₃-C₂₀ cycloalkyl, C₃-C₂₀ heterocycloalkyl,aryl, or heteroaryl; or a salt thereof.
 2. The compound of claim 1,wherein L₃ is cyclohexylene.
 3. The compound of claim 1, wherein R₅ isC₁-C₁₀ alkyl substituted with N(R_(e)R_(f)).
 4. The compound of claim 3,R₅ is


5. The compound of claim 1, R₃ is C₃-C₂₀ heterocycloalkyl substitutedwith C₁-C₁₀ alkyl, C₃-C₂₀cycloalkyl, C₃-C₂₀heterocycloalkyl, aryl, OR′,C(O)R′, COOR′, C(O)N(R′R″), SO₂R′, or C(S)N(R′R″), in which each ofR′and R″, independently, is H, C₁-C₁₀ alkyl, C₃-C₂₀ cycloalkyl, C₃-C₂₀heterocycloalkyl, aryl, or heteroaryl.
 6. A compound, wherein thecompound is


7. A compound, wherein the compound is


8. A compound of formula (I):

wherein X is —N(R_(a))—; L₁ is C₁-C₁₀ alkylene, or —C(O)—; L₂ is deletedor C₁-C₁₀ alkylene; L₃ is C₃-C₂₀ cycloalkyl; R₁ is H, C₁-C₁₀ alkyl,C₃-C₂₀ cycloalkyl, aryl, heteroaryl, halo, CN, OR_(c), COOR_(c),OC(O)R_(c), C(O)R_(c), C(O)NR_(c)R_(d), or NR_(c)R_(d); each of R₂ andR₃, independently, is H, C₁-C₁₀ alkyl, C₃-C₂₀ cycloalkyl, C₃-C₂₀heterocycloalkyl, aryl, heteroaryl, halo, CN, OR_(e), COOR_(e),OC(O)R_(e), C(O)R_(e), C(O)NR_(e)R_(f), or NR_(e)R_(f); R₄ is H; and R₅is H, C₁-C₁₀ alkyl, C₃-C₂₀ cycloalkyl, C₃-C₂₀ heterocycloalkyl, aryl, orheteroaryl; or R₄ and R₅ together are C₁-C₁₀ alkylene or C₁-C₁₀heteroalkylene; in which each of R_(a), R_(b), R_(c), R_(d), R_(e), andR_(f), independently, is H, C₁-C₁₀ alkyl, C₃-C₂₀ cycloalkyl, C₃-C₂₀heterocycloalkyl, aryl, or heteroaryl; or a salt thereof.